Figure 7.
The ROCK1/2 and MRCKα kinase genes are negative p53 targets. (A–C) Primary human keratinocytes were transfected with p53-specific siRNAs or siRNA controls and analyzed, at 48 h after transfection, for levels of ROCK1 (A), ROCK2 (B), and MRCKα (C) by real-time RT–PCR with β-actin for normalization. (D–F) Primary human keratinocytes were infected with an adenovirus expressing wild-type p53 (Ad-p53) or a GFP control (Ad-GFP) for 2 h followed by treatment with DMSO or DAPT (10 μM) for an additional 24 h. Cells were analyzed for levels of ROCK1 (D), ROCK2 (E), and MRCKα (F) mRNA expression by real-time RT–PCR as in A–C. In F, the statistical significance of the differences between Ad-GFP- and Ad-p53-infected cells with either DMSO or DAPT treatment was calculated by T-test; (*) p = 0.002; (**) p = 0.001. T-test analysis was also used to evaluate the difference in MRCKα levels after Ad-p53 infection in DMSO- versus DAPT-treated cells; (#) p = 0.03.