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Hepatic mTORC1 signaling activates ATF4 as part of its metabolic response to feeding and insulin

Byles, Vanessa, Cormerais, Yann, Kalafut, Krystle, Barrera, Victor, Hughes Hallett, James E., Sui, Shannan Ho, Asara, John M., Adams, Christopher M., Hoxhaj, Gerta, Ben-Sahra, Issam, Manning, Brendan D.·2021·Molecular Metabolism
DOI 10.1016/j.molmet.2021.101309PMC PMC8368025

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Figure 1 Feeding and insulin induce hepatic ATF4 and established gene targets by mTORC1 signaling. (A and B) Eight-week-old male mice were fasted for 12 h and refed with a high carbohydrate diet for 6 h following pretreatment with vehicle or 10 mg/kg rapamycin (n = 6/group). (A) Liver lysates were immunoblotted to assess mTORC1 signaling and ATF4, with mean ATF4 protein to tubulin quantification plotted ± SEM and normalized to the fasted group (right). (B) Liver gene expression plotted as mean ± SEM relative to the fasted group. (C) Immunoblot analysis of primary mouse hepatocytes serum-starved overnight and treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 750 nM Torin1 for the indicated time points. (D) Immunoblot analysis of primary mouse hepatocytes transfected with control (siCT) or a combination of Eif4ebp1 (4ebp1) and Eif4ebp2 (4ebp2) siRNAs, followed by overnight serum starvation and treatment for 8 h with 100 nM insulin following a 30-min pretreatment with vehicle, 20 nM rapamycin or 500 nm Torin1. (E) Immunoblot analysis of primary mouse hepatocytes treated for the indicated time points as in (C). (F) Gene expression in cells treated as in (E) plotted as mean ± SEM relative to serum-starved cells (n = 4 independent experiments). (G) Immunoblot analysis of primary human hepatocytes serum-starved overnight, and subsequently, treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 500 nM Torin1 for the indicated time points. (H) Gene expression in cells treated as in (G) plotted as mean ± SEM (n = 3) relative to the serum-starved cells. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (one-way ANOVA).
Figure 1 Feeding and insulin induce hepatic ATF4 and established gene targets by mTORC1 signaling. (A and B) Eight-week-old male mice were fasted for 12 h and refed with a high carbohydrate diet for 6 h following pretreatment with vehicle or 10 mg/kg rapamycin (n = 6/group). (A) Liver lysates were immunoblotted to assess mTORC1 signaling and ATF4, with mean ATF4 protein to tubulin quantification plotted ± SEM and normalized to the fasted group (right). (B) Liver gene expression plotted as mean ± SEM relative to the fasted group. (C) Immunoblot analysis of primary mouse hepatocytes serum-starved overnight and treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 750 nM Torin1 for the indicated time points. (D) Immunoblot analysis of primary mouse hepatocytes transfected with control (siCT) or a combination of Eif4ebp1 (4ebp1) and Eif4ebp2 (4ebp2) siRNAs, followed by overnight serum starvation and treatment for 8 h with 100 nM insulin following a 30-min pretreatment with vehicle, 20 nM rapamycin or 500 nm Torin1. (E) Immunoblot analysis of primary mouse hepatocytes treated for the indicated time points as in (C). (F) Gene expression in cells treated as in (E) plotted as mean ± SEM relative to serum-starved cells (n = 4 independent experiments). (G) Immunoblot analysis of primary human hepatocytes serum-starved overnight, and subsequently, treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 500 nM Torin1 for the indicated time points. (H) Gene expression in cells treated as in (G) plotted as mean ± SEM (n = 3) relative to the serum-starved cells. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (one-way ANOVA).CC BY-NC-ND · PubMed Central
Figure 2 ATF4 is both necessary and sufficient for insulin-mTORC1 signaling to induce select gene targets. (A) Immunoblot analysis of primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8 h. (B) Gene expression in cells treated as in (A) plotted as mean ± SEM relative to the siCT-transfected serum-starved cells (n = 3 independent experiments). (C) Immunoblot analysis of Atf4fl/fl and LAtf4KO primary mouse hepatocytes serum-starved overnight, and subsequently, treated with 100 nM insulin for 6 h following a 30-min pretreatment with vehicle (DMSO) or 20 nM rapamycin. (D) Gene expression in cells treated as in (C) plotted as mean ± SEM relative to the Atf4fl/fl serum-starved cells (n = 3). (E) Immunoblot analysis of Atf4fl/fl and LAtf4KO primary mouse hepatocytes infected with AdGFP or AdATF4 (MOI = 10) for 24 h. (F) Gene expression from a representative experiment of cells treated as in (E) plotted as mean ± SEM relative to the Atf4fl/fl AdGFP sample (n = 3). (G) Immunoblot analysis of WT primary mouse hepatocytes infected with AdGFP or AdATF4 (MOI = 10) for 24 h followed by treatment with vehicle (DMSO) or 20 nM rapamycin for 8 h. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (two-way ANOVA (B,D) or one-way ANOVA (F)).
Figure 2 ATF4 is both necessary and sufficient for insulin-mTORC1 signaling to induce select gene targets. (A) Immunoblot analysis of primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8 h. (B) Gene expression in cells treated as in (A) plotted as mean ± SEM relative to the siCT-transfected serum-starved cells (n = 3 independent experiments). (C) Immunoblot analysis of Atf4fl/fl and LAtf4KO primary mouse hepatocytes serum-starved overnight, and subsequently, treated with 100 nM insulin for 6 h following a 30-min pretreatment with vehicle (DMSO) or 20 nM rapamycin. (D) Gene expression in cells treated as in (C) plotted as mean ± SEM relative to the Atf4fl/fl serum-starved cells (n = 3). (E) Immunoblot analysis of Atf4fl/fl and LAtf4KO primary mouse hepatocytes infected with AdGFP or AdATF4 (MOI = 10) for 24 h. (F) Gene expression from a representative experiment of cells treated as in (E) plotted as mean ± SEM relative to the Atf4fl/fl AdGFP sample (n = 3). (G) Immunoblot analysis of WT primary mouse hepatocytes infected with AdGFP or AdATF4 (MOI = 10) for 24 h followed by treatment with vehicle (DMSO) or 20 nM rapamycin for 8 h. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (two-way ANOVA (B,D) or one-way ANOVA (F)).CC BY-NC-ND · PubMed Central
Figure 3 Insulin-mTORC1 signaling induces hepatic de novo nucleotide synthesis in an ATF4-independent manner. (A) Schematic of purine nucleotide highlighting carbon and nitrogen sources for de novo synthesis. (B) Protein normalized peak areas of 15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled purine nucleotide pools from primary mouse hepatocytes serum-starved overnight, and subsequently, treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 750 nM Torin1 for 8 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (C) Fractional enrichment of 15N-glutamine-amide tracing into RNA-derived purine nucleotide isotopologues in primary mouse hepatocytes serum-starved overnight, and subsequently, treated with 10 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 500 nM Torin1 for 8 h in medium containing 2 mM 15N-glutamine-amide tracer. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (D) Protein normalized peak areas of 15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled purine nucleotide pools from primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs 4 h after plating, followed by overnight serum starvation and overnight treatment with 10 nM insulin. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (E) Fractional enrichment of 15N-glutamine-amide tracing into RNA-derived purine nucleotide isotopologues in cells transfected as in (D), followed by overnight serum starvation and treatment with 10 nM insulin in medium containing 2 mM 15N-glutamine-amide tracer for 6 h. Data are plotted as mean ± SD and is representative of three independent experiments performed in triplicate. (F) Schematic of pyrimidine nucleotide highlighting carbon and nitrogen sources for de novo synthesis. (G)15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled pyrimidine nucleotide pools in primary mouse hepatocytes treated and plotted as in (B). (H) Protein normalized peak areas of 15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled pyrimidine nucleotide pools from primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs 4 h after plating, followed by overnight serum starvation and overnight treatment with 100 nM insulin. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate.
Figure 3 Insulin-mTORC1 signaling induces hepatic de novo nucleotide synthesis in an ATF4-independent manner. (A) Schematic of purine nucleotide highlighting carbon and nitrogen sources for de novo synthesis. (B) Protein normalized peak areas of 15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled purine nucleotide pools from primary mouse hepatocytes serum-starved overnight, and subsequently, treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 750 nM Torin1 for 8 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (C) Fractional enrichment of 15N-glutamine-amide tracing into RNA-derived purine nucleotide isotopologues in primary mouse hepatocytes serum-starved overnight, and subsequently, treated with 10 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 500 nM Torin1 for 8 h in medium containing 2 mM 15N-glutamine-amide tracer. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (D) Protein normalized peak areas of 15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled purine nucleotide pools from primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs 4 h after plating, followed by overnight serum starvation and overnight treatment with 10 nM insulin. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (E) Fractional enrichment of 15N-glutamine-amide tracing into RNA-derived purine nucleotide isotopologues in cells transfected as in (D), followed by overnight serum starvation and treatment with 10 nM insulin in medium containing 2 mM 15N-glutamine-amide tracer for 6 h. Data are plotted as mean ± SD and is representative of three independent experiments performed in triplicate. (F) Schematic of pyrimidine nucleotide highlighting carbon and nitrogen sources for de novo synthesis. (G)15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled pyrimidine nucleotide pools in primary mouse hepatocytes treated and plotted as in (B). (H) Protein normalized peak areas of 15N-glutamine-amide tracing (2 mM, last 30 min) into free labeled pyrimidine nucleotide pools from primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs 4 h after plating, followed by overnight serum starvation and overnight treatment with 100 nM insulin. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate.CC BY-NC-ND · PubMed Central
Figure 4 Lack of evidence for regulated serine synthesis in hepatocytes. (A) Schematic of the serine synthesis pathway. (B) Protein normalized peak areas of 15N-glutamine-amine (2 mM, last 1 h) tracing into labeled serine in primary mouse hepatocytes serum-starved overnight and subsequently treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 750 nM Torin1 for 8 h, or where indicated, cycloheximide (50 μM) was added with tracer for the last 1 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (C) Fractional enrichment of serine isotopologues from the experiment in (B). (D) Protein normalized peak areas of 15N-glutamine-amine (2 mM, last 1 h) tracing into labeled serine in primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs, followed by overnight serum starvation and overnight treatment with 10 nM insulin. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (E) Fractional enrichment of serine isotopologues from the experiment in (D), plotted as mean ± SD. (F) Protein normalized peak areas of intracellular serine from primary mouse hepatocytes following incubation in serine and/or glycine-free medium plus 2.5% dialyzed FBS for 24 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (G) Fractional enrichment of serine isotopologues in primary mouse hepatocytes treated as in (F) and labeled with 15N-glutamine-amine (2 mM, last 1 h). Data are plotted as mean ± SD and is representative of two independent experiments performed in triplicate. (H) Fractional enrichment of isotopologues from primary mouse hepatocytes following incubation in serine-rich or serine-free medium + 2.5% dialyzed FBS for 20 h with 10 mM 13C6-glucose. Data plotted as mean ± SD and is representative of two independent experiments performed in quadruplicate.
Figure 4 Lack of evidence for regulated serine synthesis in hepatocytes. (A) Schematic of the serine synthesis pathway. (B) Protein normalized peak areas of 15N-glutamine-amine (2 mM, last 1 h) tracing into labeled serine in primary mouse hepatocytes serum-starved overnight and subsequently treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 750 nM Torin1 for 8 h, or where indicated, cycloheximide (50 μM) was added with tracer for the last 1 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (C) Fractional enrichment of serine isotopologues from the experiment in (B). (D) Protein normalized peak areas of 15N-glutamine-amine (2 mM, last 1 h) tracing into labeled serine in primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs, followed by overnight serum starvation and overnight treatment with 10 nM insulin. Data are plotted as mean ± SD and are representative of two independent experiments performed in quadruplicate. (E) Fractional enrichment of serine isotopologues from the experiment in (D), plotted as mean ± SD. (F) Protein normalized peak areas of intracellular serine from primary mouse hepatocytes following incubation in serine and/or glycine-free medium plus 2.5% dialyzed FBS for 24 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (G) Fractional enrichment of serine isotopologues in primary mouse hepatocytes treated as in (F) and labeled with 15N-glutamine-amine (2 mM, last 1 h). Data are plotted as mean ± SD and is representative of two independent experiments performed in triplicate. (H) Fractional enrichment of isotopologues from primary mouse hepatocytes following incubation in serine-rich or serine-free medium + 2.5% dialyzed FBS for 20 h with 10 mM 13C6-glucose. Data plotted as mean ± SD and is representative of two independent experiments performed in quadruplicate.CC BY-NC-ND · PubMed Central
Figure 5 Insulin stimulates metabolic flux into the methionine cycle and transsulfuration pathway through ATF4. (A) Steady state metabolomic profiling from primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by 100 nM insulin stimulation for 6 h performed in quadruplicate. The heat map displays a rank order of metabolites significantly (p < 0.05) induced with insulin in the control cells, but not those with Atf4 knockdown. (B) Protein normalized peak areas of the top 6 metabolites (A) plotted as mean ± SD. (C) Schematic of the interconnections between serine and tetrahydrofolate metabolism, the methionine cycle, and the transsulfuration pathway. Metabolites found to be significantly induced by insulin in an ATF4-dependent manner in (A) are highlighted in red. (D) Gene expression analysis of primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8 h. Data are plotted as mean ± SEM relative to siCT-transfected serum-starved cells (n = 3 independent experiments). (E) Protein normalized peak areas of 3-13C-serine (400 μM, last 1 h) tracing into labeled SAM and cystathionine in primary mouse hepatocytes serum-starved overnight and subsequently treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 500 nM Torin1 for 8 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (F) Protein normalized peak areas of 3-13C-serine tracing (400 μM, last 1h) in primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (G) Protein normalized peak areas of labeled SAM from 13C5-methionine (200 μM, last 1h) tracing of cells treated as in (F). (H) Fractional enrichment of SAM isotopologues from 13C5-methionine tracing as in (G). (I) Protein normalized peak areas of labeled cystathionine from 13C5-methionine tracing as in (G). Data are plotted as mean ± SD for tracing studies and are representative of two independent experiments performed in triplicate. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (two-way ANOVA).
Figure 5 Insulin stimulates metabolic flux into the methionine cycle and transsulfuration pathway through ATF4. (A) Steady state metabolomic profiling from primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by 100 nM insulin stimulation for 6 h performed in quadruplicate. The heat map displays a rank order of metabolites significantly (p < 0.05) induced with insulin in the control cells, but not those with Atf4 knockdown. (B) Protein normalized peak areas of the top 6 metabolites (A) plotted as mean ± SD. (C) Schematic of the interconnections between serine and tetrahydrofolate metabolism, the methionine cycle, and the transsulfuration pathway. Metabolites found to be significantly induced by insulin in an ATF4-dependent manner in (A) are highlighted in red. (D) Gene expression analysis of primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8 h. Data are plotted as mean ± SEM relative to siCT-transfected serum-starved cells (n = 3 independent experiments). (E) Protein normalized peak areas of 3-13C-serine (400 μM, last 1 h) tracing into labeled SAM and cystathionine in primary mouse hepatocytes serum-starved overnight and subsequently treated with 100 nM insulin following a 30-min pretreatment with vehicle (DMSO), 20 nM rapamycin, or 500 nM Torin1 for 8 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (F) Protein normalized peak areas of 3-13C-serine tracing (400 μM, last 1h) in primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8 h. Data are plotted as mean ± SD and are representative of two independent experiments performed in triplicate. (G) Protein normalized peak areas of labeled SAM from 13C5-methionine (200 μM, last 1h) tracing of cells treated as in (F). (H) Fractional enrichment of SAM isotopologues from 13C5-methionine tracing as in (G). (I) Protein normalized peak areas of labeled cystathionine from 13C5-methionine tracing as in (G). Data are plotted as mean ± SD for tracing studies and are representative of two independent experiments performed in triplicate. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (two-way ANOVA).CC BY-NC-ND · PubMed Central
Figure 6 Characterization of feeding induced transcriptional response in LAtf4KOmice. (A,B) Eight-week-old Atf4fl/fl and LAtf4KO male mice were fasted for 12 h and refed a high carbohydrate diet for 6 h (n = 5/group). Immunoblots (A) and gene expression analysis (B) of Atf4fl/fl and LAtf4KO livers. Gene expression data are plotted as mean ± SEM relative to the Atf4fl/fl fasted group (n = 5 mice/group). (C) Principal component analysis of normalized transcripts from RNA-seq of Atf4fl/fl and LAtf4KO livers treated as in (A). (D) Heat map of normalized transcripts from RNA-seq data significantly (padj < 0.05) induced with feeding in the Atf4fl/fl, but not in the LAtf4KO livers. (E) Gene set enrichment analysis of the differentially expressed transcripts in fed LAtf4KO versus Atf4fl/fl livers. (F) Top-10 most up- and down-regulated transcripts plotted as log2 fold change in LAtf4KO versus Atf4fl/fl livers from fed mice. Asterisks indicate top hits. (G) Gene expression analysis of Atf4fl/fl and LAtf4KO livers treated and plotted as in (A). (H) Gene expression analysis of primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8h. Data are plotted as mean ± SEM relative to the siCT-transfected cells (n = 3 independent experiments). (I) Gene expression analysis of primary mouse hepatocytes from Atf4fl/fl and LAtf4KO mice infected with AdGFP or AdATF4 (MOI = 10) for 24h. Data from a representative experiment are plotted as mean ± SEM relative to the Atf4fl/fl AdGFP cells (n = 3). (J) Schematic of the Atf4 locus and proximal genes on mouse chromosome 15, along with the modified Atf4fl/fl locus. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (two-way ANOVA (B, G, I) or one-way ANOVA (H)).
Figure 6 Characterization of feeding induced transcriptional response in LAtf4KOmice. (A,B) Eight-week-old Atf4fl/fl and LAtf4KO male mice were fasted for 12 h and refed a high carbohydrate diet for 6 h (n = 5/group). Immunoblots (A) and gene expression analysis (B) of Atf4fl/fl and LAtf4KO livers. Gene expression data are plotted as mean ± SEM relative to the Atf4fl/fl fasted group (n = 5 mice/group). (C) Principal component analysis of normalized transcripts from RNA-seq of Atf4fl/fl and LAtf4KO livers treated as in (A). (D) Heat map of normalized transcripts from RNA-seq data significantly (padj < 0.05) induced with feeding in the Atf4fl/fl, but not in the LAtf4KO livers. (E) Gene set enrichment analysis of the differentially expressed transcripts in fed LAtf4KO versus Atf4fl/fl livers. (F) Top-10 most up- and down-regulated transcripts plotted as log2 fold change in LAtf4KO versus Atf4fl/fl livers from fed mice. Asterisks indicate top hits. (G) Gene expression analysis of Atf4fl/fl and LAtf4KO livers treated and plotted as in (A). (H) Gene expression analysis of primary mouse hepatocytes transfected with control (siCT) or Atf4-targeting siRNAs followed by overnight serum starvation and treatment with 100 nM insulin for 8h. Data are plotted as mean ± SEM relative to the siCT-transfected cells (n = 3 independent experiments). (I) Gene expression analysis of primary mouse hepatocytes from Atf4fl/fl and LAtf4KO mice infected with AdGFP or AdATF4 (MOI = 10) for 24h. Data from a representative experiment are plotted as mean ± SEM relative to the Atf4fl/fl AdGFP cells (n = 3). (J) Schematic of the Atf4 locus and proximal genes on mouse chromosome 15, along with the modified Atf4fl/fl locus. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 (two-way ANOVA (B, G, I) or one-way ANOVA (H)).CC BY-NC-ND · PubMed Central

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Metadata

Authors
Byles, V., Cormerais, Y., Kalafut, K., Barrera, V., Hughes Hallett, J. E., Sui, S. H., Asara, J. M., Adams, C. M., Hoxhaj, G., Ben-Sahra, I., & Manning, B. D.
ORCID
Venue
Molecular Metabolism
Publisher
Elsevier BV
Published
2021-11
Volume
53
Pages
101309
ISSN
2212-8778
DOI
10.1016/j.molmet.2021.101309
PMC
PMC8368025
Cited by
44
References
11
Language
en
Licence
Funders
  • National Institutes of Health
  • DOD (I01BX00976)
  • U.S. Department of Veterans Affairs
  • Joslin Diabetes Center (P01-CA120964, R01-AG060637, R01-AR071762, R35-CA197459, T32-NK007260, W81XWH-18-1-0659)
  • Zafgen

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