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. 2013 Oct 26;35(3):703–713. doi: 10.1093/carcin/bgt356

Fig. 3.

Fig. 3.

BNF inhibits the expression of cyclinD1/D3 and CDK4 and induces p21Cip1/Waf1 by modulating PI3K/AKT and MAPK signaling respectively. For A and B, MCF-7 cells were treated with PI3K inhibitor LY294, 002 for 48h. (A) Western blot analysis of the cell cycle proteins and the PI3K/AKT and ERK signaling proteins. (B) Flow cytometric analysis of cell cycle arrest. Upper panel, representative cell cycle distribution; Lower panel, quantitation of cell cycle distribution. For C, D and E, MCF-7 cells were preincubated with MEK inhibitor PD98059 (10 µM) for 1h and then treated with 10 µM BNF for 48h. (C) Western blot analysis of the cell cycle proteins, AKT and ERK phosporylation and (D) qPCR analysis of p21Cip1/Waf1 mRNA level. (E) Flow cytometric analysis of cell cycle arrest. Left panel, representative cell cycle distribution; Right panel, quantitation of cell cycle distribution. The data represent the mean ± SEM from four separate experiments. *P < 0.05, **P < 0.01 and ***P < 0.001, versus DMSO-treated cells; $, P < 0.05 and $$, P < 0.01, versus PD98059-treated cells; #, P < 0.05, ##, P < 0.01 and ###, P < 0.001, versus BNF-treated cells.