Figure 2
From: TGF-β upregulates the translation of USP15 via the PI3K/AKT pathway to promote p53 stability
USP15 knockdown by shRNA disrupts the p53 stability. (a) USP15 knockdown by shRNA destabilizes p53. USP15 knockdown of HEK293 cells were carried out by the different shRNAs. Cell lysates were prepared and probed for USP15, p53, p21 and actin. (b) Overexpression of the shRNA-insensitive USP15 cDNA increases the p53 stability in USP15-knockdowned U2OS cells. (c) USP15 overexpression stabilizes p53. Twenty-four hours after transfection with the indicated constructs, HEK293 cells were incubated with cycloheximide (100 μg/ml) for the indicated times. Total cell lysates were prepared and probed for p53, Flag and actin. (d) Quantification of c. The results are shown as a percentage of the p53/actin ratios in the absence of cycloheximide and are averaged from three independent experiments (means±s.d.). (e) USP15 knockdown accelerates the p53 degradation. HEK293 cells with USP15 knockdown by shRNA were cultured in the presence of cycloheximide (100 μg/ml) for the indicated time. Crude protein extracts were prepared and probed for USP15, p53 and actin. (f) Quantification of e. The results are averaged from three independent experiments (means±s.d.).
