Selective knockdown of Bnip3FL rescues cell death induced by DCA. (A) Cell viability of Panc-1 cells expressing Bnip3Δex3 in the absence or presence of 1–5 mM DCA for 24 h. Representative epifluorescence images of cells stained with vital dyes calcein AM and ethidium homodimer-1 to visualize live (green) and dead (red) cells, respectively, are shown. Bars, 40 µm. (B) Histogram for quantitative data shown in A. Data were obtained from at least n = 3–4 independent experiments counting ≥500 cells from n = 3 glass coverslips for each condition tested. (C–E) ROS (C) and cell viability (D and E) of Panc-1 cells treated with 5 mM DCA for 24 h in the absence or presence of shRNA-Bnip3FL during hypoxia. Representative epifluorescence images of cells stained with vital dyes calcein AM and ethidium homodimer to visualize live (green) and dead (red) cells, respectively, are shown. ROS production was monitored by DHE staining. Bars, 40 µm. (E) Histogram for quantitative data shown in D. Data were obtained from at least n = 3–4 independent experiments counting ≥500 cells from n = 3 glass coverslips for each condition tested. Data are expressed as mean ± SD (error bars). *, statistically different from CTRL.