Extended Data Fig. 7: Esrrg pre-mRNA splicing reshapes muscle metabolic pattern.
From: Kdm2a inhibition in skeletal muscle improves metabolic flexibility in obesity
(a-i) Using the scAAVrh.74.MHCK7.Hesrrg particles (defined as Kdm2aSKO-shEsrrg mice), skeletal muscle-specific Esrrg knockdown model was generated. Injection of male mice with scAAVs carrying shCTR was used as controls (defined as Kdm2afl/fl-shCTR, Kdm2aSKO-shCTR). In Kdm2afl/fl-shCTR, Kdm2aSKO-shCTR and Kdm2aSKO-shEsrrg mice, (a) the representative staining of Ta myofibers one month post-administration. Quantification of GFP+ myofibers (right) (n = 4 per group). GFP (green), Laminin (red) and DAPI (blue), Scale bars: 50 μm. (b) Western blot analysis of Esrrg and Pgc1a (n = 4 per group). Protein quantitative analysis (right). (c) The core body temperature monitoring under 4°C (n = 6 per group). (d) The random core body temperature at RT (n = 6 per group). (e) Representative image of Gas tissues (n = 4 per group). (f) mtDNA copy number analysis (n = 6 per group). (g) Western blot analysis of Sdhb, Ndufs1, Ndufs3 (n = 4 per group). Protein quantitative analysis (right). (h) Western blot analysis of p-ACC/ACC and Cpt1a levels (n = 4 per group). Protein quantitative analysis (right). (i) TG and T-CHO levels in Gas (left) or serum (right) (n = 4 per group). (j) Western blot analysis of Esrrg in WT mice Gas between RT and 4°C (n = 4 per group). Protein quantitative analysis (below). (k) Western blot analysis of Esrrg in WT mice Gas between ND and HFD (n = 4 per group). Protein quantitative analysis (below). (l) Western blot analysis of Esrrg in C2C12 myotubes between 37°C and 4°C (n = 4 per group). Protein quantitative analysis (below). (m) Western blot analysis of Esrrg in C2C12 myotubes after PA stimulation (n = 4 per group). Protein quantitative analysis (below). (n) Western blot analysis of ESRRG expression in BMI≤24 group, 24 <BMI <28 group and patients with BMI≥28 group, (n = 3 per group). Protein quantitative analysis (right). (o) KDM2A and ESRRG correlation analysis in human Ta tissues (n = 37). Data were expressed as mean ± SEM. Statistical significance was assessed by one-way ANOVA (a), two-way ANOVA (c) followed with Bonferroni’s multiple comparisons test, two-sided Student’s t-test (b, d, f-n) or two-sided Spearman’s correlation (o).
