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. 2014 Sep 10;289(45):31296–31309. doi: 10.1074/jbc.M114.565796

FIGURE 3.

FIGURE 3.

Depletion of K-Ras, RalA, and RalB regulates p21 expression levels in a p53-dependent manner. A549 (A) and H460 (C) cells were transfected with nontargeting (NT), K-Ras, RalA, or RalB siRNAs alone or in combination with p53 siRNA for 48 h. Cells were harvested for Western blot analysis as described under “Experimental Procedures.” A549 (B) and H460 (D) cells were transfected with p21 promoter-reporter constructs followed by transfection with the indicated siRNAs. Cells were harvested and analyzed for p21 promoter activity as described under “Experimental Procedures.” E, Calu-1 cells were transfected with the indicated siRNAs and processed for Western blot analysis as described under “Experimental Procedures.” Extra lanes on p53 and p21 blots were from a control lysate (Panc-1 cells) used as a control for the Western blot analysis. Data are representative of three independent experiments except for Calu-1 cell experiments, which were done twice.