Fig. 2: Induction of mitochondrial oxidative stress enhances sensitivity to complex I inhibition.

a Mitochondrial ROS were measured by MitoSOX staining and flow cytometry in the indicated cells grown for 60 hr in DMSO or 10 nM IACS-010759. The experiment was repeated independently three times and representative results were shown. b Quantification of mitochondrial ROS (MFI comparison) in sensitive (PATC66/108) and resistant (PATC124/148) groups after 60 hr treatment with 10 nM IACS-010759. Three biologically independent replicates per cell line. Data represent mean ± S.D between the sensitive group (2 cell lines) and resistant group (2 cell lines). c–f Cells were transfected with the hydrogen peroxide indicator pCS2+MLS-Hyper7 (mitochondrial matrix-targeted) (c) and pCS2 + HyPer7-NES (cytoplasm-targeted) (e), and treated with 10 nM IACS-010759 at the indicated times. Mitochondria were defined by staining with an antibody against the mitochondrial outer membrane protein TOMM20. Nuclei were stained with Hoechst 33578. Scale bar, 10μm. d Quantification of fluorescence intensity in (c). f Quantification of fluorescence intensity in (e). For (d) and (f), at least 50 cells with positive green fluorescence from three biologically independent replicates were calculated for each group. g Cell death was detected by propidium iodide staining and flow cytometry in PATC124 cells treated with DMSO or 10 nM IACS-010759 for 2 days, in the presence or absence of the mitochondrial-ROS inducer MitoPQ (10 μM). Data represent mean ± S.D of three biologically independent replicates. h Cell death in PATC66 cells treated with DMSO or 10 nM IACS-010759 for 3 days, in the presence or absence of the 1 μM mitochondrial ROS scavenger, MitoQ. Data represent mean ± S.D of three biologically independent replicates. i, j Mitochondrial ROS (i) and cell viability (j) were detected by flow cytometry in PATC124 cells infected with control sgRNA (sgCTRL) or sgRNA targeting SOD2 (sgSOD2) following treatment with10 nM IACS-010759 for 3 days. Data represent mean ± S.D of three biologically independent replicates. Statistical analysis by two-tailed Students’ unpaired t test with significance indicated (b, d, f, g–j). Source data are provided as a Source Data file.