Fig. 6
From: Sestrins are evolutionarily conserved mediators of exercise benefits
Sestrin is required for maximal oxidative capacity of muscle mitochondria. a Myoblasts isolated from WT and TKO mice were differentiated into myotubes. Light microscopy examination of myotube cultures after indicated days of differentiation. Scale bar, 200 µm. b–e Immunoblotting (b, c) and densitometry (d, e; values relative to WT myotube levels) of indicated proteins in WT and TKO myoblasts before (left) and after (right) myotube differentiation. (f) Phalloidin (green), mitotracker (red) and DAPI (blue) staining of WT and TKO myotubes. Scale bar, 50 µm. g Oxygen consumption rate (OCR) of WT and TKO myotubes, normalized by corresponding baseline OCR, after treatment with oligomycin, carbonyl cyanide-4-phenylhydrazone (FCCP) and rotenone/antimycin A. h Indicated mitochondrial OCR was calculated from (g), normalized by non-mitochondrial OCR. Biologically independent replicates: n = 3 (d, e), n = 4, 3 (g, h; WT and TKO, respectively). Error bars, s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 or ns, non-significant from a two-tailed student’s t test. Molecular weight markers are indicated in kDa.
