Extended Data Fig. 7: Regulation of Gdf15 by PGC-1α in cardiomyocytes.
From: Cardiac adaptation to endurance exercise training requires suppression of GDF15 via PGC-1α
a, Immunoblot for FLAG, PGC-1α, and tubulin from LPS853 liposarcoma cell lysates after infection with Ad.FLAG–PGC-1α at the indicated multiplicity of infection (MOI) for 72 h (n = 1 immunoblot per condition; experiment performed once without a separate biological replicate experiment). b, ChIP–qPCR analysis of FLAG–PGC-1α enrichment relative to IgG control at the promoters of Esrra and Fndc5 in NRVMs infected with Ad.FLAG–PGC-1α at MOI 100 for 72 h (n = 3 technical replicates from one chromatin IP, experiment independently repeated once). c, UCSC genome browser snapshots showing PGC-1α ChIP–seq peaks at the promoters of ESRRA and GDF15 in HEK293 cells from the indicated published study. d, UCSC genome browser snapshots showing PGC-1α ChIP–seq peaks at the promoters of Esrra and Gdf15 from mouse brown adipose tissue after 4 h cold exposure, from the indicated published study. e, Gene set enrichment analysis (GSEA) of the integrated stress response (ISR) gene set using transcriptomics from NRVMs treated with Ad.GFP or Ad.PGC-1α. Gene sets are shown if FDR < 0.05. Experiment performed in one biological cohort without a replication cohort. f, GSEA of ISR gene set from transcriptomic comparisons of mouse hearts WT_Sed, KO_Sed, WT_Ex and KO_Ex mice. Gene sets are shown if FDR < 0.05. n = 5 mice/group. Experiment performed in one biological cohort without a replication cohort. g, mRNA expression of ISR-related genes (Ppargc1a, Bnip3, Map1lc3b) in NRVMs treated with Ad.GFP or Ad.PGC-1α with or without tunicamycin and/or 2BAct (n = 3 biologically independent wells per group, experiment independently repeated once in a separate biological replication cohort). Data in (b) and (g) are shown as mean ± s.e.m. Statistical significance in (b) and (g) was assessed using unpaired two-tailed t-tests (b) or two-sided one-way ANOVA followed by Sidak’s multiple comparisons test (g). No statistical tests were performed in (a), (c), or (d). All GSEA results shown in (e-f) passed FDR < 0.05. Measurements were performed once per sample in all panels. Significance symbols: *P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; N.S., not significant. n refers to biologically independent replicates except for (b) where n refers to technical replicates as stated above. Exact P values: b, Esrra: 0.023 (primer pair 3), 0.026 (primer pair 5), 0.026 (primer pair 8); Fndc5: 0.014 (primer pair 6). g, Ppargc1a: 0.77 (Ad.GFP vs. Ad.GFP+Tunicamycin), 0.18 (Ad.GFP+Tunicamycin+2BAct), 0.99 (Ad.PGC-1α vs. Ad.PGC-1α+Tunicamycin), 0.69 (Ad.PGC-1α vs. Ad.PGC-1α+Tunicamycin+2BAct); Bnip3: 0.0000025 (Ad.GFP vs. Ad.GFP+Tunicamycin), 0.0020 (Ad.GFP+Tunicamycin+2BAct), 0.0085 (Ad.PGC-1α vs. Ad.PGC-1α+Tunicamycin), 0.050 (Ad.PGC-1α vs. Ad.PGC-1α+Tunicamycin+2BAct); Map1lc3b: 0.00051 (Ad.GFP vs. Ad.GFP+Tunicamycin), 0.084 (Ad.GFP+Tunicamycin+2BAct), 0.0096 (Ad.PGC-1α vs. Ad.PGC-1α+Tunicamycin), 0.73 (Ad.PGC-1α vs. Ad.PGC 1α+Tunicamycin+2BAct). Mouse strain and age: data from mouse tissues in (f) was derived from 10 week old male mice (age relative to initiation of the experiment) of the indicated strains (WT vs. KO as defined above) and n per group.
