Fig. 2

Arnidiol induces apoptosis in human breast cancer cells. For A-D, MDA-MB-231 cells were treated with various concentrations of Arn for 48 h or with Arn (60 μM) for different time intervals as indicated. a and b Apoptosis was determined by Annexin V-FITC/PI staining and flow cytometry (mean ± SD for 3 independent experiments; ^**P < 0.01 or ^***P < 0.001 compared with control). c and d The total cellular extract, cytosol and mitochondrial fractions were prepared and subjected to western blot using antibodies against total PRAP, cleaved PARP (C-PARP), cleaved caspase-3 (C-Caspase-3), cytochrome c (Cyto C) and Bax. GAPDH and COX IV were used as loading controls. For E-G, MCF-7, Eca109, SMMC-7721 and A549 cells were treated with Arn (60 μM) for 48 h. e Apoptosis was determined by Annexin V-FITC/PI staining and flow cytometry (mean ± SD for 3 independent experiments; ^***P < 0.001 compared with control). f and g The total cellular extract, cytosol and mitochondrial fractions were prepared and subjected to western blot using antibodies against total PRAP, cleaved PARP (C-PARP), cleaved caspase-3 (C-Caspase-3) and cytochrome c (Cyto C). GAPDH and COX IV were used as loading controls