Fig. 1

DNA-damaging chemotherapeutics increase the metabolism of HCT116 cells. A–F p53 wild-type (HCT^wt) and isogenic p53 null (HCT^Δp53) HCT116 cells were exposed to 10 µM irinotecan (Iri) for 24 h. A The oxygen consumption of cells was assessed using a Clarke electrode. B Cell fractionations were prepared and the expression of indicated proteins in equal amounts of mitochondrial extracts was analyzed by Western blot. TOM40 was used to control mitochondrial protein loading. The relative expression of electron transport chain (ETC) complex subunits to complex V was quantified by densitometry using ImageJ and is depicted in C. D MTT assays were performed, and corresponding cell numbers counted in parallel (refer to Fig. S1D) to calculate the MTT turnover/cell number. E The accumulation of reactive oxygen species (ROS) in cells was determined using flow cytometry. F The mRNA expression of indicated genes was assessed by qPCR. G, H HCT^wt cells were exposed to 10 µM Iri, 1 µM hydroxyurea (HU), 1 µM doxorubicin (Doxo), 0.1 mM TTFA, 0.1 µM rotenone (Rot), or 0.1 mg/ml chloramphenicol (Chlor) as single agents and in combination for 24 h. G The oxygen consumption of cells was assessed using a Clarke electrode. H The expression of indicated proteins within mitochondrial extracts was analyzed by Western blot. A The average of 4 individual experiments ± SEM. C–E, G The average of 3 individual experiments ± SEM. F, H The average of /is representative for 2 individual experiments ± SEM. Statistics for this figure: *p < 0.05; **p < 0.01; ***p < 0.001