Increases in mitochondrial ROS associated with mitochondrial membrane potential (ΔΨm) decreases in PEOL-treated cells.

HCT116 and HCT8 cells were treated with PEOL for 24 h, and then incubated whether with MitoSOX Red or TMRM as described in Materials and Methods. Representative histograms are shown in parallel with bar graphs showing the relative TMRM mean relative fluorescence intensity (A) and MitoSOX fluorescence (C) as measured by flow cytometry. Cytoplasmic cytochrome c was determined by western blotting as described in Materials and methods (B). Data represent means ± SEM (n = 3). *,**,*** represent p < 0.05, p < 0.01, p < 0.001 respectively as compared to HCT116 untreated cells. ‡, ‡‡, ‡‡‡ represent p < 0.05, p < 0.01, p < 0.001 respectively as compared to HCT8 untreated cells. p values were obtained by one-way ANOVA, followed by Fisher’s LSD test.