Fig. 4: Knockdown of Macrod1 decreases the metabolic flux from glucose into the TCA cycle.
From: MacroD1 sustains mitochondrial integrity and oxidative metabolism
A C2C12 cells transfected with either siMacroD1 or a scrambled siRNA (siMock) were incubated in the presence of uniformly stable isotope labelled (U-13C) glucose, and targeted metabolome analysis, monitoring the faith of glucose into different downstream metabolites were performed. Breakdown scheme of 13C glucose via glycolysis and TCA cycle; isotope labeled carbon atoms are depicted in red and non labelled ones in white. B Fractional labelling of lactate and pyruvate with M0, M1, etc. indicating the number of labeled carbon atoms, respectively. Data is shown as mean ± SD from 3 biological replicates. Statistical analyses were performed using a two-tailed student t-test (*, p < 0.05; **, p < 0.005; ***, p < 0.0005). C Fractional labelling of the indicated metabolites with M0, M1, etc. indicating the number of labeled carbon atoms, respectively. Data is shown as mean ± SD from 3 biological replicates. Statistical analyses were performed using a two-tailed student t-test (*, p < 0.05; **, p < 0.005; ***, p < 0.0005). D Two days after the knockdown of Macrod1, the oxygen consumption rate (OCR) of C2C12 cells was determined using a Seahorse analyzer, with addition of DMSO (as control) or oligomycin (OM), FCCP, or rotenone and antimycin (Rot/AM). Whole OCR curves are shown on the left, while comparisons between basal OCR (no treatment) and maximal OCR (post FCCP treatment) are shown on the right. Data is shown as mean ± SD from 3 biological replicates. Statistical analyses were performed using a two-tailed student t-test (*, p < 0.05; **, p < 0.005; ***, p < 0.0005). E Two days after knockdown of Macrod1 C2C12 cells were subjected to either glucose or glutamine starvation, and cell viability was assessed via flow cytometry; Data is shown as mean ± SD from 3 biological replicates. Statistical analyses were performed using a two-tailed student t-test (*, p < 0.05; **, p < 0.005; ***, p < 0.0005). Source data are provided as a Source Data file.
