Extended Data Fig. 4: Analysis of mitochondrial activity in SIRT5-deficient human myotubes.
From: SIRT5 safeguards against primate skeletal muscle ageing via desuccinylation of TBK1
a. Snapshot showing the transcript levels of SIRT1-7 gene in SIRT5+/+ and SIRT5−/− hMyotubes. b. Left, representative images of MyHC-positive myotubes in SIRT5+/+ and SIRT5−/− hMyotubes. Scale bars, 150 μm. Right, the differentiation efficiency of hMyotube was quantified. c. Western blotting analysis of the expression levels of oxidative respiratory chain complex proteins in SIRT5+/+ and SIRT5−/− hMyotubes. GAPDH was used as loading control. The representative results from one of three independent experiments. d. Left, analysis of mitochondria superoxide levels in SIRT5+/+ and SIRT5−/− hMyotubes by using MitoSOXTM Red probe. Scale bars, 100 μm. Right, positively stained area was quantified. e. Western blotting analysis of 4-HNE modified protein and SOD1 protein levels in SIRT5+/+ and SIRT5−/− hMyotubes. GAPDH was used as loading control. Two-tailed student’s t-test was used for statistical analysis and data are presented as the mean ± s.e.m. n = 3 biological replicates in b–e. P values are annotated in the figures.
