Figure 4From: Rad9 modulates the P21WAF1pathway by direct association with p53Interaction between hRad9 and p53. (A) Endogenous association of hRad9 and p53. Immunoprecipitation and Western blot analysis were performed using lysates of 293 cells. Anti-c-kit antibody was used as a negative control. (B) Immunoprecipitation using cell lysates of 293 transfectants with FLAG-tagged wild-type or phosphorylation-defective RAD9 plasmids. Each cell lysate was immunoprecipitated with anti-FLAG antibody and immunoblotted with anti-p53 (top) or anti-FLAG (bottom) antibody. HC, IgG heavy chain of the anti-FLAG antibody. (C) The time course experiment for the accumulation of phosphorylated p53 at Ser-15 after UV. 293 cells were exposed to UV at 20 J/m2 and harvested 0 to 72 hr after treatment as indicated. A Western blot analysis was performed using anti-phosphorylated p53 (Ser-15) antibody. Relative expression of phosphorylated p53 to whole p53 is shown. Data represent the means ± SD of three independent experiments. (D) The time course experiment of hRad9/p53 complex formation after exposure to UV. 293 cells were exposed to UV at 20 J/m2 and harvested 0 to 72 hr after treatment. Top, immunoprecipitation with anti-p53 antibody and Western blot analysis using anti-Rad9 antibody; middle, immunoprecipitation with anti-Rad9 antibody and Western blot with anti-p53 antibody; bottom, immunoprecipitation with anti-Rad9 antibody and Western blot with anti-phosphorylated p53 (Ser-15) antibody.Back to article page