Fig. 2.

In vitro effects of BER on induction of apoptosis (a) and reduction of ratios of Bcl-2/Bax proteins (b) in A549 cells by BER with its synergistic anticancer agents. a Induction of apoptosis in A549 cells by treatment for 48 h with BER at concentrations of 0 (0.1% DMSO vehicle as the control), 10, and 20 μM. The treated cells were stained with Hoechst 33258 and the apoptotic morphological changes in the nuclear chromatin were observed under a fluorescent microscope as described in the “Materials and methods” section. b Reduction of Bcl-2/Bax proteins in A549 cells by treatment for 48 h with BER at the concentrations of 10–60 μM (B10, B20, B40, B60) in the absence or presence of trichostatin A (TSA20) at 20 μg/L. Bay at 5 μM (BAY5, the inhibitor of NF-κB) and celecoxib at 20 μM (S20, the inhibitor of cyclooxygenase-2) were used as the positive control. The protein expressions of Bcl-2 and Bax were analyzed by Western blotting as described in the “Materials and methods” section. Anti-ß-Actin antibody was used as a sample loading control. The ratio of Bcl-2 and Bax, (the ratio of relative density of each band normalized to β-actin), shown as mean ± SD (Bar) is relative to that of 0 (0.1% DMSO vehicle) as the control (100%). For one experiment, 3 assays were carried out and only one set of gels is shown. Values with different letters (a–e) differ significantly (p < 0.05); e/s represents the significant synergistic effect (p < 0.05) compared with the treatment with its individual compound alone (e/s: B20+TSA20, p < 0.001, two-way ANOVA)