Apoptin induced pyroptosis through the ROS pathway. (A) Ad-vp3 and Ad-vt caused mitochondria aggregation towards the nucleus. (B) Red fluorescence changes to green fluorescence, measured by fluorescence microscopy following JC-1 staining. The ratio of red fluorescence to green fluorescence decreased. (C) Measurement of ROS levels. (D) The expression of cleaved-GSDME, BAX, and cytochrome c in HCT116 cells was analyzed by flow cytometry in the presence or absence of GSH. (E) Detection of the expression of bax, cytochrome c, Tom20 by western blotting. Data are shown as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001)