The DOI matches its Crossref record; 45 of the 45 references it cites were found in a registry or an index; 6 of the 34 sources its claims cite were checked for support.
Alterations of EHD1/EHD4 Protein Levels Interfere with L1/NgCAM Endocytosis in Neurons and Disrupt Axonal Targeting
Open accessAbstract
Axon growth is regulated by many proteins, including adhesion molecules, which need to be trafficked correctly to axons. The adhesion molecule L1/neuron–glia cell adhesion molecule (NgCAM) travels to axons via an endocytosis-dependent pathway (transcytosis), traversing somatodendritic endosomes. The Eps15 homology domain (EHD) family proteins (EHD1–EHD4) play important roles in endosomal recycling and possibly in endocytosis. We investigated whether EHD1 regulates L1/NgCAM trafficking in neurons. Both short hairpin-mediated downregulation and overexpression of EHD1 led to dendritic mistargeting of NgCAM. Downregulation of EHD1 showed increased endosomal accumulation of NgCAM, whereas, surprisingly, overexpression of EHD1 led to impairment of L1/NgCAM internalization in neurons but not in fibroblasts. Transferrin internalization, however, was unaffected. At longer overexpression times of EHD1, NgCAM endocytosis returned to normal, suggesting rapid upregulation of compensatory endocytic pathways. EHD1 is capable of hetero-oligomerization, and an endogenous complex of EHD1 and EHD4 was identified previously. We therefore tested whether short-term overexpression of other EHD family members showed a similar endocytosis defect. Expression of EHD4, but not of EHD3, also caused a defect in L1/NgCAM endocytosis. Oligomerization of EHD1 was required to cause NgCAM endocytosis defects, and simultaneous expression of EHD1 and EHD4 rescued NgCAM endocytosis. Therefore, balanced levels of EHD1–EHD4 are important for NgCAM endocytosis in neurons. Our data suggest that EHD1 plays roles in both endosomal recycling and a specialized endocytosis pathway in neurons used by NgCAM. We propose that EHD1 and EHD4 act as hetero-oligomeric complexes in this pathway.
Overview
AI snapshot
Written from the paper’s full text.
TL;DR
EHD1 and EHD4 levels are dosage-sensitive regulators of L1/NgCAM endocytosis in neurons, and altering either protein disrupts NgCAM axonal targeting. The data support a model in which EHD1/EHD4 hetero-oligomers, with balanced stoichiometry, act in a neuron-specific endocytosis pathway for NgCAM.
Key findings
- Overexpression of wild-type EHD1 or dominant-negative EHD1G429R reduced NgCAM axonal polarity; control neurons showed NgCAM 5–6-fold enriched on axons versus dendrites, while EHD1/EHD1G429R co-expression decreased the axon/dendrite polarity index.
- EHD1 knockdown increased dendritic surface NgCAM: control dendrite intensity was 756 AFU ± 35 SEM, versus 1233 AFU ± 44 SEM for shEHD1#1 (p<0.0001) and 921 AFU ± 35 SEM for shEHD1#2 (p<0.05).
- EHD1 or EHD1G429R overexpression impaired NgCAM endocytosis in neurons, but not in COS or NRK cells; transferrin internalization was unaffected, although EHD1G429R slowed transferrin recycling.
- EHD4 overexpression, but not EHD3, impaired NgCAM endocytosis and reduced NgCAM axonal polarity; endogenous L1 endocytosis in neurons was inhibited by EHD4 after 1 day (43% of control) but not after 3 days (94% of control).
- Oligomerization-defective EHD1V203P did not impair NgCAM endocytosis or polarity, and co-expression of EHD1 with EHD4 rescued NgCAM endocytosis and axonal polarity (A/D PI 5.0 ± 0.20 in EHD1+EHD4 cells versus 5.5 ± 0.25 in GFP controls).
Methods
- Primary embryonic day 18 rat hippocampal neurons were cultured and transfected at DIV 9–12 or nucleofected before plating; COS, NRK, and PC12 cells were also used.
- NgCAM/L1 endocytosis was measured with an acid-strip antibody uptake assay, and surface polarity was quantified as an axon/dendrite polarity index from fluorescence intensities.
- Quantification used 15–20 cells per condition for polarity/endosomal NgCAM and 150–200 cells per condition for percentage of endocytosing cells, across at least three experiments, with statistical testing in SPSS.
Limitations
- The paper states that an indirect effect of EHD1/EHD4 overexpression on NgCAM endocytosis cannot be completely ruled out, and that technical constraints prevented testing endogenous L1 endocytosis in more mature cultures.
- The paper notes that the anti-EHD1 antibody recognized multiple EHD family members on Western blots, that specific anti-EHD1/EHD4 antibodies showed only faint neuronal staining, and that the significance of neuronal nuclear EHD staining is unknown.
- Evident from the text: EHD1 knockdown achieved only 40–60% reduction, overexpression and knockdown produced distinct phenotypes, and compensatory endocytic pathways restored NgCAM/L1 endocytosis after 3–4 days, so acute versus long-term effects differ.
Model-written summary. Check it against the paper before relying on it.
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Claims the paper makes
56 Own result · 31 Own interpretation · 9 Own method · 8 Not checked yet · 6 No full text of the sources · 4 Partly supported · 3 Sources not found · 2 No basis given · 1 Supported · accuracy not yet measured
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Endocytosed NgCAM partially overlapped with cytoplasmic puncta of EHD1 and EHD4 staining (Fig. 8B), in agreement with the endosomal localization of EHD1 and EHD4 observed in fibroblasts (Grant and Caplan, 2008).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Co-expression of EHD1 rescues the endocytosis and polarity defects of EHD4
What the sources say
- Partly supportedcites Grant and Caplan, 2008 sourceExperimental — accuracy not yet measured
The section on functional roles in endocytic recycling reports that EHD4 colocalizes with early-endosome markers, while the structural-insights section describes EHD1 on tubular and punctate endosomes. The cited paper does not establish that these observations were made in fibroblasts. The NgCAM overlap is presented as the citing paper’s own Fig. 8B observation, not as a finding attributed to Grant and Caplan. (LLM, conf=0.82)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Partly supported
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Three of the four shRNA plasmids gave consistent downregulation of endogenous EHD staining in NRK cells.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Downregulation of EHD1 diminishes axonal polarization of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The levels of another EHD family member, EHD4, on the other hand, were not decreased by shEHD1 plasmids in NRK cells (data not shown).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Downregulation of EHD1 diminishes axonal polarization of NgCAM
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Consistent with the published literature, we found that steady-state loading of Tfn in cells co-expressing either EHD1 or EHD1G429R was the same as in untransfected controls (Suppl. Fig.4A and B “t0”)”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Expression of EHD1G429R, but not wild type EHD1, leads to defective recycling of transferrin
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Quantification of endogenous EHD1 in NRK cells revealed that expression of shEHD1#1 or shEHD1#2 plasmids resulted in an average of 40–60% decrease of endogenous EHD1 levels (Supplementary Figure 1).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Downregulation of EHD1 diminishes axonal polarization of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Knock-down of EHD1 using two different shEHD1 plasmids increased somatodendritic endosomal accumulation of NgCAM (Fig. 2F bottom panel, and Fig. 2G).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Up- and downregulation of EHD1 levels lead to different effects on endosomal accumulation of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: The shRNA-mediated downregulation of EHD1 is specific to EHD1 and does not affect other EHD family members (EHD2, EHD3) in neurons.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Overexpression of EHD3 did not cause impairment of NgCAM endocytosis (Fig. 4B, E).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > EHD4 affects NgCAM endocytosis and polarity
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In GFP-expressing control cells, NgCAM was 5 – 6-fold enriched on the axon compared to dendrites (Fig. 1D).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD1 or dominant-negative EHD1 diminishes axonal polarization of NgCAM
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In contrast, when EHD1 or EHD1G429R were expressed, NgCAM labeling of somatic endosomes was greatly decreased (Fig. 2B–D) compared to GFP-control cells.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Up- and downregulation of EHD1 levels lead to different effects on endosomal accumulation of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Oligomerization of EHD1 was required to cause NgCAM endocytosis defects”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Abstract
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: EHD1 and EHD4 form an endogenous hetero-oligomeric complex (or EHD1 is capable of hetero-oligomerization and an endogenous complex of EHD1 and EHD4 exists).
- Probably not a substantial claim. 26
- It may state two separate claims as one. 39
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“EHD4 was identified as an NGF-induced protein in PC12 cells, and it mediates Trk receptor endocytosis via a novel, neurotrophin-stimulated macroendocytosis pathway (Valdez et al., 2005; Valdez et al., 2007).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Partly supportedcites Valdez et al., 2005 sourceExperimental — accuracy not yet measured
The section ‘Pincher mediates TrkA internalization and endosomal Erk5 signaling in neurons’ reports that reducing Pincher inhibits NGF-induced TrkA internalization in PC12 cells. The paper also describes a novel pathway of Pincher-mediated Trk macroendocytosis. Our copy does not identify Pincher as EHD4 or state that EHD4 was identified as an NGF-induced protein in PC12 cells. (LLM, conf=0.79)
- Partly supportedcites Valdez et al., 2007 sourceExperimental — accuracy not yet measured
The Discussion (page unavailable) says that Pincher mediates Trk macroendocytosis, and the results report that a Pincher mutant blocks NGF-stimulated Trk internalization in PC12 cells. Our copy does not identify Pincher as EHD4 or say that EHD4 was identified as an NGF-induced protein in PC12 cells, so those parts of the jointly cited claim remain unsupported by this source. (LLM, conf=0.84)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“In neurons, EHD4 (Shao et al., 2002) has been implicated in endocytosis, rather than (or in addition to) recycling (Sharma et al., 2008).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Partly supportedcites Shao et al., 2002 sourceExperimental — accuracy not yet measured
The Results identify Pincher as most closely related to human EHD4 and show that it enhances NGF-induced TrkA endocytosis in PC12 cells. The Discussion says Pincher is expressed in neurons but describes its role in neuronal endosomal signaling as a candidate, not a demonstrated finding in neurons. (LLM, conf=0.84)
- Not checked — no full text of the sourcecites Sharma et al., 2008 source
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“EHD family members dimerize with themselves and with other members of the EHD family and form higher oligomeric structures which are crucial to their function (Grant and Caplan, 2008).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > EHD1 oligomerization is necessary for inhibition of NgCAM endocytosis
What the sources say
- Partly supportedcites Grant and Caplan, 2008 sourceExperimental — accuracy not yet measured
The “Oligomerization, tubulation and nucleotide hydrolysis” section describes EHD2 dimerization and assembly into higher-order rings, and says EHD oligomerization has functional significance. But the claim overgeneralizes hetero-oligomerization across the family: the paper says only EHD1–EHD4 interaction had been documented in vivo and that EHD2 appears not to hetero-oligomerize with other EHDs in vivo. It also does not establish that higher-order structures are crucial to every family member’s function. (LLM, conf=0.88)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Partly supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“EHD proteins also associate with pre-existing tubular membranes (Jovic et al., 2009).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Supportedcites Jovic et al., 2009 source
The section “EHD1 Associates with Pre-existing Tubular Membranes” reports that, in vivo, EHD proteins associate at least in part with pre-existing tubular membranes. The claim does not imply that this is their only role. (LLM, conf=0.94)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Supported
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The fluorescence intensity ranges for the two myc-tagged constructs was not different and NgCAM A/D PI remained high even at high expression levels of myc-EHD3 (data not shown).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > EHD4 affects NgCAM endocytosis and polarity
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Strikingly, despite the continued presence of overexpressed EHD1, NgCAM endocytosis after four days of overexpression was not statistically different from that of GFP control cells, whereas it was statistically significant after 18 hours of expression (Suppl. Figure 3).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Loss-of-function phenotype of EHD1 in neurons?
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In contrast, neurons co-expressing EHD1G429R showed an increased presence of NgCAM on the somatodendritic surface (Fig. 1C), resulting in a significant decrease of A/D PI (Fig. 1D).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD1 or dominant-negative EHD1 diminishes axonal polarization of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Similarly to NgCAM in mature primary neurons, L1 uptake was inhibited in PC12 cells highly overexpressing EHD4 (marked with asterisks), but not in cells expressing GFP or low levels of EHD4 (Supplementary Figure 5).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD4 impairs endocytosis of endogenous L1
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In both of these non-neuronal cells, NgCAM uptake was easily observed in control cells expressing GFP as well as in cells co-expressing EHD1 or EHD1G429R (Fig.3 A–C) and levels of endocytosed NgCAM were not statistically different between control cells and EHD1-expressing cells (Figure 3D).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD1 in non-neuronal cells does not inhibit NgCAM endocytosis
plainn = 2- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The axon/dendrite polarity index (A/D PI) for NgCAM surface localization was not changed by co-expression of EHD3 compared to GFP-expressing control cells, but co-expression of EHD4 led to a significant decrease in NgCAM axon/dendrite polarity (Fig. 4F), similar to that found for EHD1 overexpression.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > EHD4 affects NgCAM endocytosis and polarity
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Cells co-expressing EHD1 showed on average less than half of NgCAM soma fluorescence of GFP cells, whereas EHD1G429R was reduced to 65% of control.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Up- and downregulation of EHD1 levels lead to different effects on endosomal accumulation of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Quantifying the percentage of transfected cells which showed NgCAM endocytosis revealed that both EHD1 and EHD1G429R showed a significant decrease in the percentage of cells showing endocytosis (Fig. 2E).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Up- and downregulation of EHD1 levels lead to different effects on endosomal accumulation of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Surprisingly, we find that knock-down of EHD1 showed a different phenotype from EHD1 and EHD1G429R overexpression and reduced endocytosis was not apparent after four days of shEHD1 expression.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Loss-of-function phenotype of EHD1 in neurons?
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Rests on an assumption: The shRNA-mediated downregulation of EHD1 is specific to EHD1 and does not affect other EHD family members (EHD2, EHD3) in neurons.
In the paper
“As reported previously (Park et al., 2004), overexpression of EHD1 induced many short dendritic protrusions resembling filopodia and spines.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD1 or dominant-negative EHD1 diminishes axonal polarization of NgCAM
What the sources say
- Not checked — no full text of the sourcecites Park et al., 2004 source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“We noted that cells still showing NgCAM endocytosis usually expressed lower levels of EHD1, whereas endocytosis was not or barely detectable in cells with high overexpression levels.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Up- and downregulation of EHD1 levels lead to different effects on endosomal accumulation of NgCAM
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Endogenous L1 endocytosis was significantly inhibited by EHD4 overexpression after 1 day (Figure 5A,C; 43% of control levels), but not after 3 days of overexpression (Figure 5B,C; 94% of control levels).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD4 impairs endocytosis of endogenous L1
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In glial cells, short EHD-positive tubules were frequently detected (Fig.6 E), similar to staining reported for other non-neuronal cells.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Often, EHD-positive compartments could also be detected in axonal growth cones (Fig.6 D). These compartments were mostly round.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
hedged- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In addition to the punctate staining in the cytoplasm, the nucleus was also stained in neuronal cultures in many instances (Fig.6 A).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“It is striking that low level co-expression of EHD1 and EHD4 recruits them both to the plasma membrane whereas EHD1 and EHD3 are recruited onto endosomes when co-expressed.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > A Model: A role in NgCAM endocytosis for EHD1/EHD4 oligomers
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In addition, we found that transferrin recycling is disrupted by the dominant-negative EHD1G429R mutant, but not by wild type EHD1.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > Cargo-specificity of EHD1/EHD4 pathway
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Our work now shows that NgCAM endocytosis was not perturbed by EHD1 overexpression in non-neuronal cells, but strongly inhibited in neurons, uncovering an unexpected cell type-specificity for EHD1 in endocytosis.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > A cell type-specific phenotype for EHD1 in NgCAM endocytosis
boosteddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Affinity-purified anti-EHD4 antibody did not stain the nucleus of neurons, but staining was found prominently on punctate structures in the soma and dendrites (Fig.6 H, H’).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In the case of the EHD1 and EHD4 overexpression phenotype, we also found that compensatory endosomal pathways were upregulated after 3–4 days and L1/NgCAM endocytosis was no longer significantly impaired.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > Dosage-sensitivity of EHD family members
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Our results show that NgCAM endocytosis is highly dosage-sensitive to the expression of EHD1 and EHD4, but not of EHD3.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > Dosage-sensitivity of EHD family members
boosteddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“No nuclear staining was observed with the same antibody and staining protocol in fibroblast cell lines in which only the presumptive endosomal staining was visible (data not shown).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In neurons, tubules were not prominent and only occasionally could short tubules be found.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“EHD3 co-expression also could not substitute for EHD1 and did not rescue the endocytosis defect caused by EHD4 overexpression (Fig. 8D).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Co-expression of EHD1 rescues the endocytosis and polarity defects of EHD4
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
None of the sources cited for this claim could be matched to a paper in Crossref, so there is nothing to check it against.
In the paper
“Our data and that of others demonstrate that transferrin endocytosis is not disturbed by EHD1 overexpression, nor is endocytosis of AMPA receptors (Braun et al., 2005; Caplan et al., 2002; Lin et al., 2001; Park et al., 2004).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > Cargo-specificity of EHD1/EHD4 pathway
What the sources say
- Not matchedcites Braun et al., 2005
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- Not matchedcites Caplan et al., 2002
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- Not matchedcites Lin et al., 2001
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
boosted- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Not matched
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Line scans across the soma showed coinciding intensity peaks at the position of the plasma membrane for EHD1 and EHD4, but coinciding intensity peaks were not apparent for EHD3 (Suppl. Fig.6 C–E).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Co-expression of EHD1 rescues the endocytosis and polarity defects of EHD4
plaincorrelational- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“We noted that co-expression of EHD1 and EHD4 led to increased membrane-near staining for both markers whereas co-expression of EHD1 and EHD3 was overwhelmingly found on internal endosomes with no obvious membrane-near staining (Suppl. Fig.6 A,B).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Co-expression of EHD1 rescues the endocytosis and polarity defects of EHD4
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“We again see strong nuclear staining with the anti-EHD1 antibody (Fig.6 F) as well as punctate staining of presumptive endosomes throughout the soma, dendrites and axons (Fig.6F’).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“placing the EHD1-dependent defect in the endocytic (uptake per se) or endosomal (post-uptake) pathway.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD1 or dominant-negative EHD1 diminishes axonal polarization of NgCAM
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Our data suggest that EHD1 plays roles both in endosomal recycling and in a specialized endocytosis pathway in neurons used by NgCAM.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Abstract
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It may state two separate claims as one.
In the paper
“EHD2, on the other hand, appears to regulate endocytosis (Guilherme et al., 2004).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not matchedcites Guilherme et al., 2004
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
hedgedconditional- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Not matched
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Tfn recycling, but not steady-state endocytosis, was therefore impaired by EHD1G429R, but not by wild type EHD1.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Expression of EHD1G429R, but not wild type EHD1, leads to defective recycling of transferrin
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
In the paper
“L1 is involved in axonal pathfinding and guidance, axonal branching, and myelination (Maness and Schachner, 2007).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not matchedcites Maness and Schachner, 2007
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
plaincorrelational- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
- Not matched
In the paper
“In non-polarized cells, EHD1 (Rme1), EHD3, and EHD4 (pincher) function in endosomes and regulate trafficking through early and recycling endosomes (Grant and Caplan, 2008).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not checked yetcites Grant and Caplan, 2008 source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Similarly to NgCAM endocytosis therefore, L1 endocytosis was inhibited at 18 hours of EHD4 expression, but mostly restored after three days of expression.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Overexpression of EHD4 impairs endocytosis of endogenous L1
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“Other cargos, such as transferrin (Tfn) receptors, also endocytose and traverse somatodendritic endosomes, but are not ultimately sorted to axons (Park et al., 2004; Yap et al., 2008b).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not checked — no full text of the sourcecites Park et al., 2004 source
- Not matchedcites Yap et al., 2008b
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“we conclude that NgCAM: 1) either does not endocytose efficiently in the first place, or 2) it undergoes endocytosis followed by rapid recycling from a pre-early endosome (pre-EE) compartment that is membrane-near and very small in size, so that it would not appear as distinct puncta by immunofluorescence.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Up- and downregulation of EHD1 levels lead to different effects on endosomal accumulation of NgCAM
hedgeddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“Endocytosis correlates with growth cone advance on L1 substrate (Kamiguchi and Yoshihara, 2001).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not checked yetcites Kamiguchi and Yoshihara, 2001 source
Checked before — its earlier verdict is held back until a new check runs.
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Not checked yet
In the paper
“Structural studies of EHD2 suggest that oligomers of EHD2 bind to lipids and deform them into tubules, initiating budding (Daumke et al., 2007).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not checked — no full text of the sourcecites Daumke et al., 2007 source
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“As before, we checked the expression levels of myc-EHD1V203P compared to myc-EHD1 as well as GFP-EHD1V203P compared to GFP-EHD1 and found them to be in the same range (data not shown).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > EHD1 oligomerization is necessary for inhibition of NgCAM endocytosis
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“The axon/dendrite polarity index (A/D PI) is a very sensitive read-out of NgCAM/L1 polarization and even small effects due to partial downregulation can be detected.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Materials and Methods > Polarity index (PI)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: fails — The paper may give no reason why it holds.
Probably not a substantial claim. The paper may give no reason why it holds. It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“The L1 cytoplasmic tail binds a number of intracellular partners (Dickson et al., 2002; Kamiguchi et al., 1998; Maness and Schachner, 2007; Nagaraj and Hortsch, 2006), including the clathrin adaptor AP2.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not checked yetcites Dickson et al., 2002 source
Checked before — its earlier verdict is held back until a new check runs.
- Not checked — no full text of the sourcecites Kamiguchi et al., 1998 source
- Not matchedcites Maness and Schachner, 2007
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Not checked yet
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“For quantifying axonal polarity or somatic endosomal NgCAM, 15–20 cells were quantified per conditions in each of minimally 3 experiments.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Materials and Methods > Quantification and statistical analysis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Expression levels of transfected proteins were compared for differences in expression levels. If necessary, plasmid concentrations were adjusted to equalize expression levels.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Materials and Methods > Quantification and statistical analysis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“We argue that this observation suggests the hypothesis that transferrin recycling might be mediated by EHD1 homodimers/oligomers in neurons.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > Cargo-specificity of EHD1/EHD4 pathway
hedgedconditional- Asserted: fails — The authors may not put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The authors may not put it forward as true. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Interestingly, the balance of EHD1 and EHD4 levels appears crucial since overexpression of either one inhibits endocytosis.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > A Model: A role in NgCAM endocytosis for EHD1/EHD4 oligomers
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“The fixation conditions used do not introduce holes into the overwhelming majority of cells.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Materials and Methods > Immunocytochemistry
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: fails — The paper may give no reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Pre-absorption of the antibody with the immunizing peptide abolished the staining (data not shown).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“For determining the percentage of cells endocytosing NgCAM, 150–200 cells per condition and experiment were counted in minimally three separate experiments.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Materials and Methods > Quantification and statistical analysis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
In the paper
“Proteins participating in multimeric functional complexes are often sensitive to expression levels since the balance of subunits is crucial for correctly assembling the oligomeric structure (Papp et al., 2003).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > EHD1 oligomerization is necessary for inhibition of NgCAM endocytosis
What the sources say
- Not checked — no full text of the sourcecites Papp et al., 2003 source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Nuclear staining was competed by incubation with the immunizing peptide (data not shown).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Results > Localization of EHD1 and EHD4 in neurons
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Cells were excluded from scoring if their morphology is abnormal, indicating poor health.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Materials and Methods > Quantification and statistical analysis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“The EHD proteins have a highly conserved domain structure. The N-terminal G domain is required for ATP binding and membrane recruitment, the central region for dimerization/oligomerization, and the C-terminal EH domain for binding to NPF motifs in a number of proteins (Grant and Caplan, 2008).”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Introduction
What the sources say
- Not checked yetcites Grant and Caplan, 2008 source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Our work raises the intriguing possibility that in neurons EHD1 and EHD4 play a role directly in NgCAM endocytosis at the plasma membrane.”
§ Alterations of EHD1/EHD4 protein levels interfere with L1/NgCAM endocytosis in neurons and disrupt axonal targeting > Discussion > A Model: A role in NgCAM endocytosis for EHD1/EHD4 oligomers
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
The paper relies on this without support.
EHD1 and EHD4 form an endogenous hetero-oligomeric complex (or EHD1 is capable of hetero-oligomerization and an endogenous complex of EHD1 and EHD4 exists).
“EHD1 is capable of hetero-oligomerization, and an endogenous complex of EHD1 and EHD4 was identified previously.”
The paper relies on this without support.
The shRNA-mediated downregulation of EHD1 is specific to EHD1 and does not affect other EHD family members (EHD2, EHD3) in neurons.
“To specifically knockdown the EHD1 protein expression, we used short hairpin RNA targeting the rat EHD1 sequences obtained from Super Array Bioscience.”
The paper relies on this without support.
The distribution of EHD proteins has not been investigated in neurons.
“The distribution of EHD proteins for both endogenous and exogenous protein has been characterized in several non-neuronal cell types (for examples see (Blume et al., 2007; George et al., 2007), but has not been investigated in neurons.”
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Metadata
- Authors
- Yap, C. C. C., Lasiecka, Z. M., Caplan, S., & Winckler, B.
- Affiliations
- University of Virginia (US)
- University of Nebraska Medical Center (US)
- Corresponding author
- Winckler, Bettina
- Venue
- The Journal of Neuroscience
- Publisher
- Society for Neuroscience
- Published
- 2010-05-12
- Volume
- 30
- Issue
- 19
- Pages
- 6646-6657
- ISSN
- 0270-6474 (ISSN-L)
- 1529-2401
- PMC
- PMC2905050
- OpenAlex
- W1978427724
- Semantic Scholar
- 0304bef310b4273f760da96d4a59993c4c0ec981
- Cited by
- 44
- References
- 45
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2022: 1 · 2020: 3 · 2018: 1 · 2017: 1 · 2016: 7 · 2015: 5 · 2014: 1 · 2013: 3 · 2012: 6
- FWCI
- 1.96
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM074876, R01GM086913)
- National Center for Research Resources (P20RR018759)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Nerve injury and regeneration
- Keywords
- EHD1, EHD4, endocytosis, axon targeting, transcytosis, endosomal recycling, hetero-oligomerization, axon growth, neurons, transferrin internalization
- MeSH terms
- Animals, Axons, Cells, Cultured, Chlorocebus aethiops, Endocytosis, Fibroblasts, Hippocampus, Nerve Tissue Proteins, Neurons, Endosomes, Time Factors, Transferrin, Signal Transduction, PC12 Cells, COS Cells, Vesicular Transport Proteins, Neural Cell Adhesion Molecule L1, Cell Enlargement, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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