Fig. 2: Nebivolol inhibits mitochondrial ATP synthase.

HCT116 and MDA-MB-231 cells were treated during 3 h with 1 µM nebivolol (NEB; red dots and bars), oligomycin (OL; gray dots and bars) or left untreated (CRL; closed dots or bars). a Left panel, kinetic representation of the production of ATP in digitonin-permeabilized cells. The inhibition of ATP synthase activity was accomplished by the addition of 30 μM OL. Right panel, histograms show the ATP synthetic activity. Six replicates of six (HCT116, **p = 0.001 and **p = 0.00001) and three (MDA-MB-231, **p = 0.004 and **p = 0001) different biological samples are shown. b Glycolytic flux measured by the initial rates of lactate production. Three replicates of three different biological HCT116 (*p = 0.01 and 0.01) and MDA-MB-231 (*p = 0.04 and p = 0.01) samples are shown. c Plots and histograms of TMRM⁺ stained cells to assess the mitochondria membrane potential (ΔΨm). FCCP (orange) collapses ΔΨm. Three replicates of three different biological HCT116 (*p = 0.01 and 0.01) and MDA-MB-231 (*p = 0.05 and 0.02) samples are shown. See Supplementary Fig. 7a for gating strategy. d Histograms show ROS production in HCT116 (*p = 0.02) and MDA-MB-231 (*p = 0.05) cells treated (red) or not (black) for 3 h with 1 µM nebivolol. Three replicates of three different biological samples are shown. See Supplementary Fig. 7b for gating strategy. e Representative western blots of four different biological samples of the expression of mitochondrial proteins from complex I (NDUFA9), complex II (SDHB), complex III (Core 2), complex IV (COX IV), complex V (β-F1-ATPase) and IF1 (*p = 0.011 and 0.02), Hsp60 and GAPDH and tubulin as loading controls. Two different samples of control (CRL; closed bars) and nebivolol-treated (NEB; red bars) cells are shown. Bars indicate the mean ± SEM of the indicated samples. *p < 0.05 and **p < 0.01 when compared to CRL by two-sided Student’s t test. See also Supplementary Fig. 2. Source data are provided as a Source Data file.