Figure 3: Mitochondrial aspartate synthesis by PC is essential for Sdhb−/− cell proliferation and survival. | Nature Communications

Figure 3: Mitochondrial aspartate synthesis by PC is essential for Sdhb−/− cell proliferation and survival.

From: Loss of succinate dehydrogenase activity results in dependency on pyruvate carboxylation for cellular anabolism

Figure 3

(a) Successful knockdown of PC expression in both wild-type (WT) and Sdhb−/− cells assessed by quantitative RT–PCR (n=3, error bars are s.e.m), results in (b) inhibition of proliferation and decrease in survival only in Sdhb−/− (KO c8) cells, whereas scrambled (scr) siRNA shows no effect (n=3, error bars are s.e.m). (c,d) Glutamate/aspartate antiporter SLC25A13 protein expression in (d) Sdhb−/− cells and (e) SDHB-mutated PCC consistent with increased requirement for export of aspartate from the mitochondria. Scale bar, 100 μm. (e) Sdhb−/− imCC (KO c6 and c8) demonstrate increased uptake of aspartate when incubated with 0.1 mM 13C4-aspartate for 48 h compared with wild-type cells, consistent with an increased requirement for aspartate in SDH-deficient cells due to dysfunctional synthesis (n=3, error bars are s.d.). *P=0.05, Kruskal–Wallis test. (f) Labelled aspartate is metabolized more rapidly into cellular metabolites such as succinate in Sdhb−/− cells (n=3, error bars are s.d.). MID, mass isotopomer distribution KO, Sdhb knockout.

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