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. Author manuscript; available in PMC: 2020 Jun 6.
Published in final edited form as: Mol Cell. 2019 Apr 22;74(5):877–890.e6. doi: 10.1016/j.molcel.2019.03.031

Figure 1: Limited glucose availability promotes mitochondrial oxygen consumption, Respiratory Chain Supercomplexes and Cristae density.

Figure 1:

U2OS cells grown in glucose or galactose for 48h. Oxygen consumption analysis of (A) intact cells, or (B) isolated mitochondria, using Pyruvate/Malate or Succinate as specific substrates respectively. (C) Mitochondrial enzymatic activities of CI, CIV, CII, CI+III and CII+III normalized to citrate synthase (CS) levels. (D) Upper panel; Immunodetection of the indicated proteins representing CI, CIII, CIV and CII after BNGE of digitonin-solubilized mitochondria from glucose or galactose (48h). Right; Bands were quantified using ImageJ software and fold induction under galactose conditions is represented. Lower panel; Superposition profiles of the different supercomplexes and free complexes using GelEval software. (E) Complex I in-gel activity of digitonin-solubilized mitochondria from 48h glucose or galactose cultured cells. (F) Mitochondrial cristae morphology, abundance and density assayed by electron microscopy (EM) in glucose and galactose-grown U2OS cells at 24h or 48h. (G), Mean fluorescence corresponding to mitochondrial content in cells cultured in glucose or galactose measured by flow cytometry. SDHA was used as a loading control. Immunoblots shown are representative of >3 independent experiments and all other experiments are represented as mean ± s.e.m., n>3. Asterisks denote *p<0.05 or **p<0.01. For two comparisons a two-tailed t-test was used, for multiple comparisons, one-way ANOVA with Bonferroni post-test was applied. gluc/g, glucose. Galac/G, galactose.