Skip to main content
. 2011 Nov 15;30(24):4921–4930. doi: 10.1038/emboj.2011.419

Figure 1.

Figure 1

Identification of USP42 as a regulator of p53 stability. (A) p53-inducible Saos-2 cells stably expressing PG13-luciferase reporter were transfected with control or a pool of USP42 siRNA oligonucleotides. Forty-eight hours post transfection, p53 was induced by treatment with 1 μg/ml doxycycline. Cells were lysed 24 h after p53 induction and luciferase activity was determined using a luciferase assay system (Promega). (B) p53-inducible Saos-2 cells were transfected with control or individual USP42 siRNA oligonucleotides. Forty-eight hours post transfection, p53 was induced by treatment with 1 μg/ml doxycycline. Total cell lysates were prepared 24 h after p53 induction. Lysate proteins were resolved by SDS–PAGE and analysed by western blotting with anti-p53 DO1 and Actin antibodies. (C) p53-inducible Saos-2 cells were transfected with control or USP42 siRNA oligonucleotides. Forty-eight hours post transfection, p53 was induced by treatment with 1 μg/ml doxycycline, then 24 h after p53 induction, cells were treated with 100 μg/ml cyclohexamide for the indicated times. Total cell lysates were resolved by SDS–PAGE and analysed by western blotting with anti-p53 and anti-Actin antibodies. Quantification of three independent experiments is shown in the graph below.