Fig. 3

BAM15 reduces OXPHOS and glycolytic capacity via ΔΨm destabilization in MDA-MB-231 and EO771 cells. Oxygen consumption rates following acute injection of varying concentrations of BAM15 in A MDA-MB-231 (Veh N=5, 1 μM BAM15 N=5, 10 μM BAM15 N=3, 20 μM BAM15 N=4) and B EO771 cells (Veh N=6 per condition). Respiration supported by malate, pyruvate, and glutamate (N-linked), or succinate (S-linked) in the presence of ADP, oligomycin, and varying concentrations of BAM15 in digitonin-permeabilized cells in C MDA-MB-231 (N=6 per condition) and D EO771 cells (N=4 per condition). ATP-linked respiration in E MDA-MB-231 and F EO771 cells (N=4 per condition). Maximal glycolytic rate in G MDA-MB-231 and H EO771 cells the presence of glucose and oligomycin (N=4 per condition for MDA-MB-231; N=5 per condition for EO771 cells) following 16-h exposure to vehicle or varying concentrations of BAM15. Respiration supported by malate, pyruvate, glutamate, and succinate in the presence of ADP, FCCP, and ascorbate/TMPD after 16-h exposure to vehicle or 20 μM BAM15 in digitonin-permeabilized I MDA-MB-231 (N=8 per condition) and J EO771 cells (N=4 per condition). Superoxide production following 16-h exposure to vehicle or 20 μM BAM15 in K MDA-MB-231 (N=4 per condition) and L EO771 cells (N =4 per condition). M Representative flow cytometry plot, and N quantification of TMRM florescence after acute exposure to varying concentrations of BAM15 or FCCP (N=6 per condition) and O, P chronic exposure to varying concentrations of BAM15 (N=5 per condition) in MDA-MB-231 cells. Q Representative flow cytometry plot, and R quantification of TMRM florescence after acute exposure to varying concentrations of BAM15 or FCCP (N=6 per condition) and S, T chronic exposure to varying concentrations of BAM15 (N=6 per condition) in EO771 cells. Data are shown as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Panels C, D, E, F, G, H, K, L, N, P, R, and T were assessed by one-way ANOVA with Tukey’s multiple comparisons. Panels A, B, I, and J were assessed by two-way repeated measures ANOVA with Sidak’s multiple comparisons. Abbreviations: OCR, oxygen consumption rate; ECAR, extra-cellular acidification rate; Dig, digitonin; ADP, adenosine 5′-diphosphate; O, oligomycin; PM, pyruvate and malate; G, glutamate; S, succinate; FCCP, Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone; TMPD, tetramethyl-p-phenylene diamine; TMRM, tetramethylrhodamine methyl ester