Fig. 6.

PGC-1α regulates BCAA gene transcription through ERRα. (a) STRING Interacting Network showing the top five interacting proteins for PGC-1α. (b) Efficiency of ESRRA silencing in primary HSMCs. (c) Expression of BCAA genes. (d) Leucine oxidation rates in HSMCs. (e) Schematic biochemical representation of the leucine oxidation assay. (f) Expression of ESRRA, PPARGC1A and target genes in Ad-PGC1A cells transfected with ESRRA siRNA. (g–i) BCAA gene expression in Ad-PGC1A cells with or without ESRRA siRNA. (j) Expression of ESRRA in skeletal muscle biopsies from individuals with NGT or type 2 diabetes. Gene expression is shown as log₂(fold change) relative to the corresponding scramble+GFP treated cells (dotted line), scr (b, c), scr + Ad-GFP (f–i) or NGT fasting (j). Statistical analysis was performed using paired t test (b, c, n = 7), two-way repeated measures ANOVA followed by Tukey’s post hoc test (d, n = 4; f–i, n = 6, except in g: DLD, n = 5), or two-way mixed-design ANOVA followed by Sidak’s post hoc test (j, n = 25). *p < 0.05, **p < 0.01 and ***p < 0.001 vs scr (b, c) or scr-Ad-GFP (f–i). ‡, siESRRA effect; §, Ad-PGC1A effect; ¶, interaction effect. Main and interaction effects symbols indicate p < 0.05 to p < 0.0001. See also ESM Fig. 4.; BT2, 3,6-dichlorobrenzo(b)thiophene-2-carboxylic acid; ESR1, oestrogen receptor 1; FC, fold change; scr, scrambled siRNA; SIRT1, sirtuin 1; TCA, tricarboxylic acid cycle