Fig. 4: Effect of mitochondrial ROS quenchers on ferroptosis.

A SK-Hep1 ρ⁺ cells were treated with 5 μM erastin or 0.1 μM RSL3 for 24 h in the presence or absence of 100 μM MitoTEMPO or 500 nM MitoQ. Cell death was determined by LDH release assay. B SK-Hep1 ρ⁺ and ρ⁰ cells were treated with 1.0 μM RSL3 in the presence or absence of MitoTEMPO for 24 h. Cell death was determined by LDH release assay. C, D In SK-Hep1 ρ⁺ treated with 5 μM erastin in the presence or absence of 100 μM MitoTEMPO or 500 nM MitoQ for 16 h, accumulation of mitochondrial ROS (C) and mitochondrial lipid peroxides (D) was determined by flow cytometry after staining with MitoSOX and MitoPerOx, respectively (right). Representative histograms are shown (left). E, F In SK-Hep1 ρ⁺ treated with 0.1 μM RSL3 in the presence or absence of 100 μM MitoTEMPO or 500 nM MitoQ for 4 h, accumulation of mitochondrial ROS (E) and mitochondrial lipid peroxides (F) was determined by flow cytometry after staining with MitoSOX and MitoPerOx, respectively (right). Representative histograms are shown (left). G, H SK-Hep1 ρ⁺ cells were treated with 5 μM erastin (G) or 0.1 μM RSL3 (H) for 24 h in the presence or absence of varying concentrations of DecylQ and MitoQ. Cell death was determined by LDH release assay. Data represent means ± SD from three independent experiments. Data were analyzed by one-way ANOVA with Tukey’s multiple comparison test (A–F) or two-tailed unpaired Student’s t test (G, H). (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant).