Fig. 5

p53 is translocated into mitochondria in response to ROS, which contributes to CTB-induced apoptosis. a SMMC-7721 cells were treated with CTB at the indicated concentrations (0, 1, 2, 4 μΜ) for 24 h. Western blot analysis of p53 expression in cytoplasm or mitochondria respectively in SMMC-7721 cells. b Cells treated with CTB (2 μM) for the indicated time periods (1, 2, 4, 8, 16, 20, 24 h). Western blot analyses of p53 accumulation in mitochondria. c Representative Fluorescence microscope imaging of SMMC-7721 cells treated with CTB (2 μΜ) for 24 h, labeled with DAPI, anti-p53 antibody and Mito-tracker Green. Scale bar: 10 μm. d SMMC-7721 cells treated with the indicated concentrations of Pifthrin-μ (10 μM), CTB (2 μM) and Pifthrin-μ (10 μM) + CTB (2 μM) for 24 h. Representative Fluorescence microscope imaging of SMMC-7721 cells labeled with DAPI, anti-p53 antibody and Mito-tracker Green. Scale bar: 10 μm. e Western blot analyses of p53 expression in mitochondria. f The opening of mPTP was detected by Rhodamine123. Scale bar: 50 μm. g Western blot analysis of Bcl-2, Bax and p53 protein levels in mitochondria, and Cyt C release from mitochondria to cytoplasm. Data are represented as mean ± SD. Significance: *P < 0.05, **P < 0.01 and ***P < 0.001 vs Control; ^#P < 0.05, ^##P < 0.01 and ^###P < 0.01 vs CTB (2 μΜ) treatment