Fig. 5
The mechanism of DC661-induced cell apoptosis. A Immunofluorescent staining of cathepsin B in HCC cells treated with DC661 (3 µM, 6 h). Representative cells showing that DC661 treatment led to leakage of lysosomal cathepsin B are shown. Scale bar, 10 μm. B Western blot showing the level of Bax and Bcl-2 in HCC cells treated with DC661 (3 µM, 6 h). Data represent mean ± SD; **P < 0.01. C Effects of DC661 treatment on mitochondrial membrane potential (ΔΨm). Cells were incubated with DC661 at 3 µM for 6 h, then stained with JC-1. Hoechst 33,258 staining indicates the nuclei. Scale bar, 20 μm. D Immunofluorescence co-staining of cytochrome c and Tom20 (indicates the mitochondria). Cells were incubated with DC661 at 3 µM for 6 h. Hoechst 33,258 staining indicates the nuclei. Scale bar, 20 μm. E, F Flow cytometric analysis and statistical data of the level of activated caspase-3 in HCC cells treated with DC661 (1 µM, 24 h). Data represent mean ± SD; ***P < 0.001. G, H Flow cytometric analysis and statistical data of cell apoptosis and necrosis in HCC cells treated with DC661 (1 µM, 24 h). Data represent mean ± SD; *P < 0.05; **P < 0.01