Effects of BER with its synergistic anticancer agents on Bcl-2/Bax protein expressions in MDA-MB-231 cells. Western blot analysis of Bcl-2 (A) and Bax (B) expressions in whole-cell lysates of MDA-MB-231 (left panel) and MDA-MB-435S (right panel) cells treated for 48 h with BER at the indicated concentrations; (C) Reduction of Bcl-2/Bax ratio in MDA-MB-231 cells by BER and its synergistic anticancer agents celecoxib and trichostatin A. The cells were treated for 48 h with the indicated concentrations of BER (B, 20 μM), celecoxib (S, 20 μM), trichostatin A (T, 50 μg/L), Ly 294002 (LY, 25 μM), and Bay (5 μM) in the absence or presence of its synergistic anticancer agents S (20 μM) and T (50 μg/L). The expressions of Bcl-2 and Bax in MDA-MB-231 were analyzed by Western blotting. In the (A) and (B), the density of the band (normalized to β-actin) shown as mean ± SD (Bar) is relative to that of 0 as the control (designated as 100%). In the (C), 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 (designated as 1.0). For one experiment, 3 assays were carried out and only one set of gels is shown. Comparison was made by two-way ANOVA followed by Bonferroni post hoc test to establish whether significant differences existed between the groups. *: P < 0.05, **, P < 0.01. Values with different letters (a-e) differ significantly (P < 0.05). e/s+ represents the significant synergistic effects (P < 0.05) compared with the treatment with its individual compound alone. Statistically significant synergistic effect on the Bcl-2/Bax ratio was observed in MDA-MB-231 cells treated with B20+S20 or B20+T50 compared with the individual B20, S20 or T50 treatment alone (B20+S20, P < 0.001, two-way ANOVA; B20+T50, P < 0.001, two-way ANOVA).