Fig. 2.

Regulation of p53 stability by USP11

(a) U2OS cells were transfected with increasing amounts of Flag-USP11. Cell lysates were prepared and analyzed by Western blotting with the indicated antibodies 24 h post-transfection. (b) p53 mRNA levels of U2OS control cells and USP11-expressing cells were measured by quantitative real-time PCR. Results are shown as mean±standard deviation (SD) of triplicate experiments and normalized to the control. (c) Half-life assay of endogenous p53 protein. U2OS cells transfected with vector or Flag-USP11 were treated with 30 μg/ml cycloheximide (CHX) and harvested at the indicated time. Protein levels were detected by Western blotting using the indicated antibodies. The values are obtained from three independent experiments and are normalized to the actin control. For each experimental condition, the signal at the start of the experiment was set to 1. CS: Cys 275 mutants to Ser