Fig. 3

BTW induces G2/M cell cycle arrest and activates mitochondrial apoptosis in PAC cells. Representative images of cell cycle distribution of PAC cells (A, B) following BTW treatment at concentrations of 0, 10, 20 and 40 μg/mL for duration of 12, 24 and 48 h, respectively. Quantification data of cell cycle distribution of PAC cells (C, D). The expression of the indicated proteins in PAC cells (E, F) was analyzed using immunoblotting after treatment with 0–40 μg/mL of BTW for 12 h. The expression levels were normalized to β-actin. Double staining of Annexin V-FITC and PI was conducted to detect apoptotic cell death in PAC cells (G, H) following BTW treatment (0–40 μg/mL) for 48 h. Quantification analysis was shown on the right panel. The expression of apoptosis-related proteins was evaluated using immunoblotting in MIA PaCa-2 (I, J) and BxPC-3 (K, L) cells following BTW treatment. β-actin was used as an internal reference for protein expression normalization. 0.5 μM of GEM was chosen as the positive control. Results are presented as mean ± S.D. of triplicate independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the control