Fig. 7

p20BAP31 induced AIF-mediated caspase-independent apoptosis. A HCT116 cells were pretreated with Z-VAD-FMK (20 μM) before transfection with p20BAP31 for 48 h, and the apoptosis rate was assessed by flow cytometry. B Quantitative analysis of apoptosis rates by flow cytometry. C Western blot analysis detected the protein expression of whole cellular, mitochondria, and nuclear AIF after transfection with p20BAP31 for 48 h. D The relative quantification of the protein levels was analyzed by ImageJ software. E Immunostaining of HCT116 cells was carried out with an anti-AIF antibody and a FITC secondary antibody (green) to localize AIF. Hoechst 33,342 staining (blue) was used to visualize the nuclei, and the merged images clearly demonstrate the nuclear localization of AIF. ***P < 0.001