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. 2007 Jun 22;104(27):11388–11393. doi: 10.1073/pnas.0609467104

Fig. 5.

Fig. 5.

p53 accumulation is inhibited by Cul7. (A) Western blot analysis of MCF-7 cells stably transfected with pRTS-1 episomal expression vector (39) encoding FlagCul7. See SI Fig. 10B for pRTS-1-Luciferase (pRTS-1-Ctrl) vector control experiment. Cul7 expression was induced by addition of 500 ng/ml DOX for 48 h. Cells were treated with etoposide (20 μM) for the indicated periods. (B) H1299 cells were transfected with plasmids encoding p53, Cul7, and FBX29. Shown is Western blot analysis of whole cell extracts (WCL) and immunoprecipitates (IP; obtained with a HA-specific antibody). (C) In vivo ubiquitination assay for p53 in H1299 cells. Cells were transfected with pMT107 [His-tagged ubiquitin (38)], pcDNA3-p53-VSV, and plasmids expressing MDM2 or FBX29 plus Cul7, as indicated. For details, see Materials and Methods. Circle with an X, background signal originating from anti-VSV antibody. Bottom lane, Western blot analysis of whole cell lysate.