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. 2012 Dec 14;5(3):929–934. doi: 10.3892/ol.2012.1077

Figure 4.

Figure 4

Effects of NOC on the expression of Bcl-2 family proteins in MDA-MB-453 cells. (A) MDA-MB-453 cells were treated with the indicated concentrations of NOC or chrysin for 24 h. Expressions of Bax, PUMA, Noxa, Bim, Bcl-2 and Bcl-XL proteins were examined by western blotting of total cell lysates and β-actin was used as the loading control. (B) MDA-MB-453 cells were transfected with 100 nM siRNA control or the siRNA duplexes against FOXO3a mRNA. Forty-eight hours after transfection, cells were treated with 4.0 μM NOC for 24 h. Expression of Bim protein was determined by western blotting of total cell lysates and β-actin was used as the loading control. (C) MDA-MB-453 cells were transfected with 100 nM siRNA control or the siRNA duplexes against Bim mRNA. Forty-eight hours after transfection, cells were treated with 4.0 μM NOC for 24 h. Expression of Bim protein was determined by western blotting of total cell lysates and β-actin was used as the loading control. (D) MDA-MB-453 cells were transfected with 100 nM siRNA control or siRNA duplexes against Bim mRNA. Forty-eight hours after transfection, the cells were treated with 4.0 μM NOC for 24 h. The apoptotic rate was analyzed by FCM using PI staining. Data shown are means ± SD (n=3). *P<0.05; **P<0.01 vs. 0.1% DMSO; #P<0.05 vs. the same concentration of NOC in combination with siRNA Bim transfection. NOC, 7-dihydroxy-8-nitrochrysin; Bcl-2, B cell lymphoma 2; Bax, Bcl-2 associated X protein; PUMA, p53 upregulated modulator of apoptosis; FCM, flow cytometry; PI, propidium iodide; SD, standard deviation; DMSO, dimethyl sulfoxide.