Student or educator? Sign up with your school email and get 6 months of Pro free.
CoChat
Sign Up
Cited by 501

The DOI matches its Crossref record; 5 of the 5 references it cites were found in a registry or an index.

Nuclear Factor of Activated T-cells (NFAT) Rescues Osteoclastogenesis in Precursors Lacking c-Fos

Matsuo, Koichi, Galson, Deborah L., Zhao, Chen, Peng, Lan, Laplace, Catherine, Wang, Kent Z.Q., Bachler, Marcus A., Amano, Hitoshi, Aburatani, Hiroyuki, Ishikawa, Hiromichi, Wagner, Erwin F.·2004·Journal of Biological Chemistry
DOI 10.1074/jbc.m313973200
Open this paper in CoChat — free account

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1074/jbc.m313973200

Cites

5 of 5 references verified0 citation errors · 0 not checkable

Statistics

Figures

Image 1
CC-BY · From the PDF
FIG. 2 c-Fos binds to the Nfatc1 promoter. A , schematic presentation of putative Fos ( open ovals ) and NFAT ( closed rectangles ) binding sites in the mouse Nfatc1 promoter (27, 28). The numbering is relative to the proximal transcriptional start site directed by promoter P1. Distal ( dis. ) and proximal ( prox. ) blocks of homology are shown in thick lines . Arrows , PCR primers. B , chromatin immunoprecipitation assay. Non-adherent Fos -/ -splenocytes and wild-type bone marrow cells were treated with or without RANKL for 20 h before cross-linking. Precipitation was performed with IgG or anti-c-Fos antibody ( 𝛂 Fos ), and the amounts of precipitated DNA relative to total input DNA were quantified by a real-time PCR for Nfatc1 ( top ) and Acp5 ( bottom ; see Fig. 3 A for PCR primers).
FIG. 2 c-Fos binds to the Nfatc1 promoter. A , schematic presentation of putative Fos ( open ovals ) and NFAT ( closed rectangles ) binding sites in the mouse Nfatc1 promoter (27, 28). The numbering is relative to the proximal transcriptional start site directed by promoter P1. Distal ( dis. ) and proximal ( prox. ) blocks of homology are shown in thick lines . Arrows , PCR primers. B , chromatin immunoprecipitation assay. Non-adherent Fos -/ -splenocytes and wild-type bone marrow cells were treated with or without RANKL for 20 h before cross-linking. Precipitation was performed with IgG or anti-c-Fos antibody ( 𝛂 Fos ), and the amounts of precipitated DNA relative to total input DNA were quantified by a real-time PCR for Nfatc1 ( top ) and Acp5 ( bottom ; see Fig. 3 A for PCR primers).CC-BY · From the PDF
FIG. 3 Identification of NFAT/AP-1 site in the Acp5 promoter. A , conserved Acp5 promoter sequences in mouse ( M ), human ( H ), and pig ( P ). Asterisks indicate nucleotides identical to the 15-bp mouse IL-2 distal NFAT/AP-1 site. Arrows indicate PCR primers for the chromatin immunoprecipitation assay in Fig. 2 B . B , EMSA using Acp5 -120 ( 5 ), IL-2 distal NFAT/AP-1 site ( I ) and AP-1 consensus ( A ) or mutant ( m ) sites for probes and competitors ( Comp. ). Nuclear extracts containing NFAT binding activity were prepared from osteoclastogenic co-culture. C , transient transfection assay in RAW264.7 cells using wild-type ( WT ) and mutant ( MUT ) mouse Acp5 promoter-luciferase constructs. The activator plasmid expressing 𝚫 NFAT ( 𝚫 ) was co-transfected. RLU , relative light units (normalized to co-transfected Renilla luciferase activity and relative to the unstimulated wild-type promoter). D , the Acp5 promoter-luciferase construct was activated by c-Fos expression vector ( Fos ) and suppressed by Fos small interfering RNA vectors ( RNAi1 , RNAi2 ) in transient co-transfection in RAW264.7 cells.
FIG. 3 Identification of NFAT/AP-1 site in the Acp5 promoter. A , conserved Acp5 promoter sequences in mouse ( M ), human ( H ), and pig ( P ). Asterisks indicate nucleotides identical to the 15-bp mouse IL-2 distal NFAT/AP-1 site. Arrows indicate PCR primers for the chromatin immunoprecipitation assay in Fig. 2 B . B , EMSA using Acp5 -120 ( 5 ), IL-2 distal NFAT/AP-1 site ( I ) and AP-1 consensus ( A ) or mutant ( m ) sites for probes and competitors ( Comp. ). Nuclear extracts containing NFAT binding activity were prepared from osteoclastogenic co-culture. C , transient transfection assay in RAW264.7 cells using wild-type ( WT ) and mutant ( MUT ) mouse Acp5 promoter-luciferase constructs. The activator plasmid expressing 𝚫 NFAT ( 𝚫 ) was co-transfected. RLU , relative light units (normalized to co-transfected Renilla luciferase activity and relative to the unstimulated wild-type promoter). D , the Acp5 promoter-luciferase construct was activated by c-Fos expression vector ( Fos ) and suppressed by Fos small interfering RNA vectors ( RNAi1 , RNAi2 ) in transient co-transfection in RAW264.7 cells.CC-BY · From the PDF
FIG. 5 Rescue of osteoclast formation with 𝚫 NFAT. A , TRAP staining of osteoclastogenic co-cultures derived from wild-type ( + / + ) splenocytes and GFP and 𝚫 NFAT virus-infected Fos -/ -splenocytes ( -/ -). Below , GFP fluorescence and TRAP stain are shown in higher magnification. The bars represent 100 𝛍 m. B , bone resorption assay. Infected cells were co-cultured with ST2-T cells on bovine bone slices for 10 days, and pit formation was examined in a double-blind study with backscattered electron imaging. The bars represent 0.75 mm. Arrows indicate bone resorption pits. C , rescue activity of 𝚫 NFAT virus relative to that of c-Fos virus. st , osteoclastogenic co-cultures using ST2-T cells. M + R , osteoblast-free cultures using soluble M-CSF and RANKL. Fos -/ -splenocytes were infected with GFP, c-Fos, or 𝚫 NFAT viruses. Calcr transcripts were quantitated by real-time PCR; TRAP-positive multinucleated cells ( MNC ) were counted, and the resorbed area (%) was measured on bone surfaces.
FIG. 5 Rescue of osteoclast formation with 𝚫 NFAT. A , TRAP staining of osteoclastogenic co-cultures derived from wild-type ( + / + ) splenocytes and GFP and 𝚫 NFAT virus-infected Fos -/ -splenocytes ( -/ -). Below , GFP fluorescence and TRAP stain are shown in higher magnification. The bars represent 100 𝛍 m. B , bone resorption assay. Infected cells were co-cultured with ST2-T cells on bovine bone slices for 10 days, and pit formation was examined in a double-blind study with backscattered electron imaging. The bars represent 0.75 mm. Arrows indicate bone resorption pits. C , rescue activity of 𝚫 NFAT virus relative to that of c-Fos virus. st , osteoclastogenic co-cultures using ST2-T cells. M + R , osteoblast-free cultures using soluble M-CSF and RANKL. Fos -/ -splenocytes were infected with GFP, c-Fos, or 𝚫 NFAT viruses. Calcr transcripts were quantitated by real-time PCR; TRAP-positive multinucleated cells ( MNC ) were counted, and the resorbed area (%) was measured on bone surfaces.CC-BY · From the PDF
FIG. 6 Rescue of osteoclast formation with 𝚫 NFAT in the absence of RANKL. A , transient transfection of 𝚫 NFAT-expression vector induces endogenous Acp5 and Calcr expression in RAW264.7 cells untreated or treated with 30 ng/ml RANKL as indicated and harvested at the days depicted. Osteoclasts generated in co-culture served as positive control ( lane 8 ) and no RNA as negative control ( lane 1 ). Transcripts were analyzed by reverse transcriptase-PCR with primers for Acp5 , Calcr (all isoforms), and glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ). The reverse transcripts-PCR products for Calcr were also detected by hybridization with an internal probe ( hyb ). B , rescue of TRAP-positive Fos -/ -cells by 𝚫 NFAT requires RANKL. M-CSF-dependent macrophages derived from wild-type and Fos -/ -splenocytes ( Sp + / + and Sp -/ -) and wild-type bone marrow cells ( Bm + / + ) were infected with the GFP 𝚫 NFAT virus and then were cultured for 3 more days in the presence or absence of RANKL. The bottom row shows expression of GFP 𝚫 NFAT fusion protein.
FIG. 6 Rescue of osteoclast formation with 𝚫 NFAT in the absence of RANKL. A , transient transfection of 𝚫 NFAT-expression vector induces endogenous Acp5 and Calcr expression in RAW264.7 cells untreated or treated with 30 ng/ml RANKL as indicated and harvested at the days depicted. Osteoclasts generated in co-culture served as positive control ( lane 8 ) and no RNA as negative control ( lane 1 ). Transcripts were analyzed by reverse transcriptase-PCR with primers for Acp5 , Calcr (all isoforms), and glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ). The reverse transcripts-PCR products for Calcr were also detected by hybridization with an internal probe ( hyb ). B , rescue of TRAP-positive Fos -/ -cells by 𝚫 NFAT requires RANKL. M-CSF-dependent macrophages derived from wild-type and Fos -/ -splenocytes ( Sp + / + and Sp -/ -) and wild-type bone marrow cells ( Bm + / + ) were infected with the GFP 𝚫 NFAT virus and then were cultured for 3 more days in the presence or absence of RANKL. The bottom row shows expression of GFP 𝚫 NFAT fusion protein.CC-BY · From the PDF

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 5 references

Research works printed but not linked: 29

  1. Chinenov, 2001, Oncogene, 20, 2438, 10.1038/sj.onc.1204385 · DOI
  2. Teitelbaum, 2003, Nat. Rev. Genet., 4, 638, 10.1038/nrg1122 · DOI
  3. Karsenty, 2002, Dev. Cell, 2, 389, 10.1016/S1534-5807(02)00157-0 · DOI
  4. Komarova, 2003, J. Biol. Chem., 278, 8286, 10.1074/jbc.M206421200 · DOI
  5. Wang, 1992, Nature, 360, 741, 10.1038/360741a0 · DOI
  6. Johnson, 1992, Cell, 71, 577, 10.1016/0092-8674(92)90592-Z · DOI
  7. Grigoriadis, 1994, Science, 266, 443, 10.1126/science.7939685 · DOI
  8. Matsuo, 2000, Nat. Genet., 24, 184, 10.1038/72855 · DOI
  9. Fleischmann, 2000, Genes Dev., 14, 2695, 10.1101/gad.187900 · DOI
  10. Takayanagi, 2002, Nature, 416, 744, 10.1038/416744a · DOI
  11. Hoey, 1995, Immunity, 2, 461, 10.1016/1074-7613(95)90027-6 · DOI
  12. Crabtree, 2002, Cell, 109, S67, 10.1016/S0092-8674(02)00699-2 · DOI · Paper page
  13. Hirotani, 2004, J. Biol. Chem., 279, 13984, 10.1074/jbc.M213067200 · DOI
  14. Lean, 2000, Bone, 27, 29, 10.1016/S8756-3282(00)00306-9 · DOI
  15. Schreiber, 1989, Nucleic Acids Res., 17, 6419, 10.1093/nar/17.15.6419 · DOI
  16. Molkentin, 1998, Cell, 93, 215, 10.1016/S0092-8674(00)81573-1 · DOI
  17. Onishi, 1996, Exp. Hematol., 24, 324
  18. Anusaksathien, 2001, J. Biol. Chem., 276, 22663, 10.1074/jbc.M007104200 · DOI
  19. Barton, 2002, Proc. Natl. Acad. Sci. U. S. A., 99, 14943, 10.1073/pnas.242594499 · DOI
  20. Weinmann, 2002, Methods (Orlando), 26, 37
  21. Lacey, 1998, Cell, 93, 165, 10.1016/S0092-8674(00)81569-X · DOI · Paper page
  22. Sherman, 1999, J. Immunol., 162, 2820, 10.4049/jimmunol.162.5.2820 · DOI
  23. Chuvpilo, 2002, Immunity, 16, 881, 10.1016/S1074-7613(02)00329-1 · DOI
  24. Zhou, 2002, J. Biol. Chem., 277, 10704, 10.1074/jbc.M107068200 · DOI
  25. Diebold, 1998, Proc. Natl. Acad. Sci. U. S. A., 95, 7915, 10.1073/pnas.95.14.7915 · DOI
  26. Luchin, 2001, J. Biol. Chem., 276, 36703, 10.1074/jbc.M106418200 · DOI
  27. Macian, 2001, Oncogene, 20, 2476, 10.1038/sj.onc.1204386 · DOI
  28. Stroud, 2003, J. Mol. Biol., 334, 1009, 10.1016/j.jmb.2003.09.065 · DOI · Paper page
  29. Kim, 2002, J. Exp. Med., 195, 201, 10.1084/jem.20011681 · DOI

Cited by

See all 501

Metadata

Authors
Matsuo, K., Galson, D. L., Zhao, C., Peng, L., Laplace, C., Wang, K. Z. Q., Bachler, M. A., Amano, H., Aburatani, H., Ishikawa, H., & Wagner, E. F.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2004-06
Volume
279
Issue
25
Pages
26475-26480
ISSN
0021-9258
DOI
10.1074/jbc.m313973200
Cited by
501
References
5
Language
en
Licence

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.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: Nuclear Factor of Activated T-cells (NFAT) Rescues Osteoclastogenesis in Precursors Lacking c-Fos. https://app.cochat.ai/papers/10.1074/jbc.m313973200
The paper
Matsuo, K., Galson, D. L., Zhao, C., Peng, L., Laplace, C., Wang, K. Z. Q., Bachler, M. A., Amano, H., Aburatani, H., Ishikawa, H., & Wagner, E. F. (2004). Nuclear Factor of Activated T-cells (NFAT) Rescues Osteoclastogenesis in Precursors Lacking c-Fos. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.m313973200

Data sources

Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.