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. 2012 Feb 16;7(2):e32267. doi: 10.1371/journal.pone.0032267

Figure 5. Opposing effect of BITC treatment on levels of Bcl-xL and Bcl-2 proteins in MDA-MB-231 cells.

Figure 5

(A) Western blotting for Bcl-xL and Bcl-2 using lysates from MDA-MB-231 cells treated with DMSO (control) or BITC (2.5 or 5 µM) for the indicated time periods. Number above band indicates change in level compared to the corresponding DMSO-treated control. (B) Western blotting for Bcl-xL and (C) quantitation of histone-associated DNA fragment release into the cytosol in MDA-MB-231 cells transiently transfected with empty vector or vector encoding for Bcl-xL and treated for 24 h with DMSO or 5 µM BITC. Results are expressed as enrichment relative to corresponding DMSO-treated control. Data represent mean ± SD (n = 3). Significantly different (P<0.05) acompared with corresponding DMSO-treated control, and bbetween empty vector transfected cells and Bcl-xL overexpressing cells by one-way ANOVA followed by Bonferroni's multiple comparison test. (D) Western blotting for Bcl-2 and (E) quantitation of histone-associated DNA fragment release into the cytosol in MDA-MB-231 cells transiently transfected with a control siRNA or the Bcl-2-targeted siRNA and treated for 24 h with DMSO or 5 µM BITC. In panel E, results shown are relative to control siRNA transfected cells treated with DMSO. Data represent mean ± SD (n = 3). Significantly different (P<0.05) acompared with corresponding DMSO-treated control, and bbetween control siRNA transfected cells and Bcl-2 siRNA transfected cells by one-way ANOVA followed by Bonferroni's multiple comparison test. (F) Western blotting for p53 and Bcl-2 using lysates from MCF-7 cells transiently transfected with a control siRNA or the p53-targeted siRNA and treated for 24 h with DMSO or 5 µM BITC. (G) Western blotting for p53 and PUMA using lysates from MCF-7 cells transiently transfected with a control siRNA or the p53-targeted siRNA and treated for 24 h with DMSO or 5 µM BITC. Number above the bands in 5F, G represents change in level relative to control siRNA transfected cells treated with DMSO. All the experiments were repeated at least twice and representative data from one such experiment are shown.