Fig. 4

CHCHD4-mediated tumour cell growth is linked to CI-regulated mTORC1 signalling and amino acid metabolism. a Schematic of proposed model of CHCHD4-regulated CI-dependent amino acid (AA) metabolism and influence on tumour cell growth. b Chart shows relative intracellular abundance of selected metabolites from metabolomics analysis in CHCHD4 (WT)-expressing cells (WT.cl1) relative to control U2OS cells. Representative of 2 experiments. ± SD. n = 5. c Chart shows relative proportions of isotopically labelled metabolites in control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1). Representative of 2 experiments. n = 5. d Western blots show levels of phosphorylated (P-), total (T-) p70S6K and puromycin labelled polypeptides in control U2OS cells and CHCHD4 (WT)- expressing cells (WT.cl1) treated as indicated for 8 h. β-Actin was used as a load control. Relative band intensities of P-p70S6K indicated. e Images of control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1, WT.cl3) cultured to confluency in the presence of glutamine (6 mM), and 3 days later following removal of glutamine (0 mM). f Western blots show levels of phosphorylated (P-) and total (T-) p70S6K, and puromycin labelled polypeptides in control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1) treated either in the absence (−) or presence (+) of 50 nM BAY 87-2243 for 24 h, supplemented with 10 mM aspartate (D) or 1 mM NAD. β-Actin was used as a load control. g Charts show relative growth of control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1) treated with 10 nM BAY 87-2243 in the absence or presence of 10 mM aspartate (D) or 1 mM NAD for 72 h. ± SD. n = 3