Fig 1: Expression levels of ETC complex subunits and PGC1-NRF1/2 signaling pathway proteins during differentiation of C2C12 myoblasts. (a) Western blot of ETC complex subunits ND1, CYTB, ATP6A1, SDHA, and COXIV in C2C12 cells during differentiation on D1, D3, and D5 after treatment with UCF101 (20 µM) or no treatment and quantification of Western blots relative to GM (n = 3). (b) Western blot of PGC1a, NRF1, NRF2, and VDAC in C2C12 myoblasts at differentiation on D1, D3, and D5 after treatment with UCF101 (20 µM) or no treatment and quantification of Western blots relative to GM (n = 3). Two-tailed unpaired Student’s t-tests were used. Statistical significance: * p = 0.05, ** p = 0.01. ND1, NADH1; SDHA, succinate dehydrogenase complex flavoprotein subunit A; COXIV, cytochrome c oxidase subunit 4; CYTB, cytochrome b; ATP6, ATP synthase F0 subunit 6.
Fig 2: mRNA and protein expression regulated by cryptotanshinone in PCOS. (a) mRNA expression of PGC1a as determined by RT-qPCR. (b) mRNA expression of NRF1 as determined by RT-qPCR. (c) mRNA expression of GPX4 as determined by RT-qPCR. (d) mRNA expression of catalase gene as determined by RT-qPCR. (e) mRNA expression of SOD1 as determined by RT-qPCR. (f) Blot images and numeric quantification of protein expression of PGC1a, NRF1, GPX4, catalase, ERK, p-ERK, UCP2, BAX, and BCL2 as determined by western blotting. *p < 0.05, compared to control; #p < 0.05 compared to IR; $p < 0.05 compared to IR+CRY; &p < 0.05 compared to IR+CRY+ferrostatin-1.
Fig 3: Upstream regulators for ALS proteins. (a) Table of upstream regulators, the number of their binding partners, their predicted activation state and score, and p-value of overlap. (b) Representative image of PPARG expression in the Betz cells of control cases (n = 12). PPARG is detected in the cytoplasm and in some cases in the nucleus. (c) Representative images of PPARG expression in the Betz cells of sporadic ALS patients (n = 8). (d) Representative images of PPARG expression in the Betz cells of ALS patients with TDP43 pathology (n = 9). (e) Representative image of PPARGC1A expression in the Betz cells of control cases (n = 12). PPARG is low but is detected in the cytoplasm. (f) Representative images of PPARGC1A expression in the Betz cells of sporadic ALS patients (n = 8). (g) Representative images of PPARGC1A expression in the Betz cells of sporadic ALS patients (n = 8). PPARGC1A expression is increased and the proteins is accumulated in the cytoplasm, z-score =2 and p = 0.05 is significant. Scale bar = 50 µm in b-g.
Fig 4: Maternal Exercise Stimulates DNA Hypomethylation of Ppargc1a Promoter(A) The ratio of a-ketoglutarate (a-KG) to 2-hydroxyglutarate (2-HG) in fetal muscle at E18.5 (n = 6).(B) Diagram showing three regions in the Ppargc1a proximal promoter.(C) 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) enrichment fold of fetal skeletal muscle in response to maternal exercise (n = 6).(D) 5mC enrichment fold in the gastrocnemius muscle of female and male M-Ctrl offspring challenged with CD or HFD (n = 6).(E and F) 5mC and 5hmC enrichment fold of offspring gastrocnemius muscle after CD/HFD challenge in response to maternal exercise (E; n = 6) and fetal muscle following apelin administration during pregnancy (F; n = 6).(G) A proposed mechanism of maternal-exercise-dependent AMPK/a-KG/Ppargc1a demethylation axis, stimulating mitochondrial biogenesis (PGC-1a1) and endurance performance.Data are mean ± SEM, and each dot represents one litter; two-sided p values by unpaired Student’s t test (A–F). See also Figure S5.
Fig 5: PPARGC1A activates slow–twitch muscle phenotype and induces muscle hypertrophy. (A–J), relative glycogen content (A), relative mRNA expression levels of glycogenolytic and glycolytic genes (B), relative enzyme activity of lactic dehydrogenase (LDH) and succinate dehydrogenase (SDH) (C), immunohistochemistry analysis of MYH1A/MYH7B (D), MYH1A/MYH7B protein content (E), relative mRNA expression levels of several fast–/slow–twitch myofiber genes (F), relative gastrocnemius muscle weight (G), H&E staining (H), and frequency distribution of fiber cross-sectional area (CSA) (I,J) in gastrocnemius with PPARGC1A overexpression or under–expression. In panels (A–C,E–G), results are shown as mean ± SEM, statistical significance of differences between means was assessed using paired t–tests. (* p < 0.05; ** p < 0.01).
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