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. Author manuscript; available in PMC: 2013 Dec 27.
Published in final edited form as: Cell Cycle. 2008 Feb 11;7(9):1205–1213. doi: 10.4161/cc.7.9.5756

Figure 4.

Figure 4

HAUSP downregulation inhibits stress-induced translocation of p53 to mitochondria. (A) Apoptosis-associated p53 phosphorylation is intact upon HAUSP downregulation. Cells pretreated with Dox were stressed with Camptothecin for 5 hrs and immunoprecipitated with DO1. Input into immunoblots was adjusted for equal amounts of immunoprecipitated p53 in stressed cells (see CM-1 lanes). (B) p53 transactivation function upon HAUSP downregula-tion. Real time qRT-PCR of cells induced with Dox for 48 hrs. Average fold induction of Noxa and Puma after treatment with Camptothecin for 4 hrs (Noxa) and 9 hrs (Puma). Whiskers indicate standard error of triplicates. (C) Left, HAUSP downregulation inhibits stress-induced p53 translocation to mitochondria. Immunoblots of purified mitochondria from cells pretreated with Dox, followed by Camptothecin stress for 4 hrs. Mitochondrial mthsp70 as loading control. Right, immunoblot from total cell lysates of stressed cells +/− Dox. PCNA as loading control. (D) HAUSP downregulation decreases p53 monoubiquitination in response to stress. Cells induced with Dox were treated with Camptothecin for 1 hr, followed by the proteosome inhibitor ALLN for 6 hrs. Input into immu-noblot was normalized for equal amounts of p53 (short exposure). Long exposure of the same blot reveals the difference in monoubiquitination.