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. 2013 Jun 4;8(6):e66275. doi: 10.1371/journal.pone.0066275

Figure 2. Rac1/Cdc42 regulates p53 expression to affect VEGF expression in MCF-7 cells.

Figure 2

(A–D) MCF-7 cells were transfected with Rac1/Cdc42 small-interfering RNA (siRac1/Cdc42) or with negative control siRNA and with the plasmid pCEFL-GST-V12Rac1 (V12Rac1), pCEFL-GST-L61Cdc42 (L61Cdc42) or pCEFL-GST-neo using lipofectamine 2000 for 48 h. Protein and RNA were then extracted and subjected to western blot analyses. Total protein was subjected to 15% SDS-PAGE; membranes were incubated with anti-Rac1, anti-Cdc42, anti-p53, anti-VEGF, or anti-β-actin antibody. (E–H) MCF-7 cells were transfected with siRac1/Cdc42 or with p53 siRNA (sip53) and either activated Rac1/Cdc42 plasmid (V12Rac1/L61Cdc42) or wild-type p53 expression plasmid (wt-p53) with lipofectamine 2000 for 48 h. Protein was then extracted and subjected to 15% SDS-PAGE; membranes were incubated with anti-p53, anti-VEGF, or anti-β-actin antibody. The data represent three independent experiments.