Fig. 3: Reduced accumulation of mitochondrial ROS and mitochondrial lipid peroxides in SK-Hep1 ρ⁰ cells.

A SK-Hep1 ρ⁺ and ρ⁰ cells were treated with 5 μM erastin in the presence or absence of 2.5 μM Fer-1 or 0.1 μM Lip-1 for 16 h and then analyzed for mitochondrial ROS by flow cytometry after staining with MitoSOX (right). Representative histograms are shown (left panel). B, C SK-Hep1 ρ⁺ and ρ⁰ cells were treated with 0.1 (B) or 1.0 μM (C) RSL3 in the presence or absence of 2.5 μM Fer-1 or 0.1 μM Lip-1 for 4 h and then analyzed for mitochondrial ROS by flow cytometry after staining with MitoSOX (right). Representative histograms are shown (left panel). D SK-Hep1 ρ⁺ cells were treated with 5 μM erastin in the presence or absence of 2.5 μM Fer-1 or 0.1 μM Lip-1 for 16 h and then analyzed for mitochondrial peroxidized lipids by flow cytometry after staining with MitoPerOx (right). Representative histograms are shown (left panel). E, F SK-Hep1 ρ⁺ and ρ⁰ cells were treated with 0.1 (E) or 1.0 μM (F) RSL3 in the presence or absence of 2.5 μM Fer-1 or 0.1 μM Lip-1 for 4 h and then analyzed for mitochondrial peroxidized lipids by flow cytometry after staining with MitoPerOx (right). Representative histograms are shown (left panel). Data represent means ± SD from three independent experiments. Data were analyzed by one-way ANOVA with Tukey’s multiple comparison test (A–F except ρ⁺ and ρ⁰ cell comparisons) or two-tailed unpaired Student’s t test (ρ⁺ and ρ⁰ comparisons in A–F). (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not significant).