Figure 5
Investigation on the apoptotic regulatory functions of Cg-Bak and Cg-Bax. (a) HeLa cells were transfected with Cg-Bak-EGFP or pEGFP-N1 empty vector and UV-irradiated or left untreated. Cell nuclei were stained with Hoechst 33342 (blue) and Alexa Fluor 633 (red) or MitoView 633 (red) to stain the cell membrane or mitochondria, respectively, as indicated in the figure. The green fluorescent signal indicated the distribution of overexpressed proteins. (b) HeLa cells were transfected with Cg-Bax-EGFP or pEGFP-N1 empty vector (green) and then UV-irradiated or left untreated. Cell nuclei were stained with Hoechst 33342 (blue). (c) Cg-Bak-EGFP and Cg-Bax-EGFP were overexpressed in HEK293T cells, which were then subjected to UV irradiation. Cytosolic and mitochondrial extracts from non-irradiated and irradiated cells were examined by western blot with anti-GFP antibody. (d) Isolated mitochondria from oyster cells were incubated with storage buffer, CaCl2 (300 μM), recombinant Cg-Bak protein (8 μM), or recombinant Cg-Bax (8 μM) at room temperature for 30 min prior to centrifugation. The distributions of cytochrome c in the supernatant (S) and precipitate (P) were then analyzed by western blot
