1. mGPDH effect occurs via the CaMKKβ/AMPK control of mitochondrial biogenesis
mGPDH effect occurs via the CaMKKβ/AMPK control of mitochondrial biogenesis A–FMitochondrial DNA (A), nuclear‐encoded OXPHOS genes (B), respirometry analysis (C), and immunoblots of mGPDH, phospho‐Thr172 AMPK (p‐AMPK), total AMPK (AMPK), phospho‐Ser79‐ACC (p‐ACC), total ACC and PGC1α, and corresponding quantifications represent mGPDH, p‐AMPK, p‐ACC, and PGC1α protein levels (D–F) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation.G–IImmunoblots of p‐AMPK, p‐ACC, and PGC1α and corresponding quantifications represent p‐AMPK, p‐ACC, and PGC1α protein levels (G), mitochondrial DNA (H), and nuclear‐encoded OXPHOS genes combined by NDUFS8, SDHb, Uqcrc1, COX5b, and ATP5a1 (I) in C2C12 myocytes transfected by mGPDH plasmid with the AMPK inhibitor compound C (CC) 24 h after differentiation.J, KNAD+/NADH ratio (J) and immunoprecipitation analysis for PGC1α acetyl‐lysine (Ac‐Lys) level (K) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation.L–PImmunoblot of c‐myc and myogenin (L) and corresponding quantifications represent c‐myc and myogenin protein levels (M), representative images of MyHC immunofluorescence (N), fusion index (O), and the distribution of nuclei per myotube (P) in C2C12 myocytes transfected with mGPDH plasmid with the AMPK inhibitor CC at 24 h (L, M) or 72 h (N–P) after differentiation.QImmunoblots of p‐AMPK, p‐ACC, PGC1α, and myogenin in C2C12 myocytes transfected with mGPDH plasmid with the CaMKKβ inhibitor STO‐609 at 24 h after differentiation. Quantifications represent p‐AMPK, p‐ACC, PGC1α, and myogenin protein levels.RImmunoblots of p‐AMPK and p‐ACC in C2C12 myocytes transfected with mGPDH plasmid with the Ca2+ chelator BAPTA‐AM at 24 h after differentiation. Quantifications represent p‐AMPK and p‐ACC protein levels.Data information: Data are presented as the mean ± s.e.m. Scale bars represent 50 μm in panel (N). In panels (A, B, D–M, Q, and R), n = 3; in panel (C), n = 10; in panels (N–P), n = 15. *P < 0.05, **P < 0.01, ***P < 0.001, n.s.: not significant. Unpaired t‐test was used in panels (A–C, E, F, and J); one‐way ANOVA with Tukey's comparison test was used in panels (G–I, M, O, Q, and R); and the Kolmogorov–Smirnov test was used in panel (P).Source data are available online for this figure.
Xiufei Liu et al. EMBO Mol Med. 2018;emmm
© as stated in the article, figure or figure legend