Figure 5

ATF2 localization induces mitochondrial membrane damage, mitochondrial cytochrome c release, and caspase activation in B16F10 cells. (A) B16F10 cells stably transfected with scrambled or ATF2 shRNA were incubated with either PTX or CCCP (50 M, 1 h). The cells were stained with JC-1 dye and observed the mitochondrial membrane potential by fluorescence microscopy. Green fluorescence indicates JC-1 stained mitochondrial plasma with a low JC-1 concentration (monomer) and depolarized mitochondrial membrane potential; red fluorescence indicates JC-1 stained mitochondrial plasma with a high JC-1 concentration (polymer)(Scale bar: 20 μm). (B) B16F10 cells stably transfected with scrambled or ATF2 shRNA were treated as indicated above. JC-1 flow cytometry analysis was performed and the represented diagrams of flow cytometry were presented. B16F10 cells stably transfected with scrambled or ATF2 shRNA were treated as indicated. The cells then were subjected to immunofluorescently stain with cytochrome c (Scale bar: 20 μm) (C) and immunoblot analysis for the mitochondrial and cytoplasmic extracts with the indicated antibodies (D).