FIG. 9.

Effect of a nonacetylated p53 mutant on transcriptional repression. (A) Dose-response analysis of wild-type p53 (p53wt) and mutant p53-9KR in Saos2 cells with the cyclin B2 and Mdm2 reporters. Error bars indicate standard deviations. (B) Levels of expression of wild-type p53 and p53-9KR in panel A are shown in Western blots. (C) Immunoprecipitation of overexpressed wild-type p53 and mutant p53-9KR from transfected H1299 cells. Western blot analysis was performed with anti-p53 and anti-YA antibodies. WCE, whole-cell extracts; Ctl, control. (D) ChIP analysis of NF-Y and p53 in NIH 3T3 cells transfected with wild-type p53, mutant p53-9KR, and control vectors, together with the cyclin B2-luciferase construct, before and after adriamycin (ADR) treatment for 8 h. PCRs with the transfected templates are shown in the right panels, and those with the control endogenous cyclin B2 gene are shown in the left panels. Two sets of PCRs are shown. Note that fewer PCR cycles are required for the transfected templates.