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. Author manuscript; available in PMC: 2012 May 22.
Published in final edited form as: Cancer Res. 2010 Dec 6;71(3):884–894. doi: 10.1158/0008-5472.CAN-10-2518

Figure 4. 14-3-3σ antagonizes COP1-mediated p53 degradation.

Figure 4

A, 293T cells were cotransfected with the indicated plasmids. Lysates were immunoblotted with the indicated antibodies.

B, H1299 cells were cotransfected with the indicated plasmids. Cells were treated with MG132 before harvesting. The polyubiquitinated p53 were immunoprecipitated with anti-p53 and immunoblotted with anti-HA. Lysates were also immunoblotted with the indicated antibodies.

C, p53−/−MDM2−/− MEF cells were cotransfected with the indicated plasmids. Cells were treated with MG132 before harvesting. Polyubiquitination of p53 was detected by immunoprecipitation with anti-p53 and immunoblotting with anti-ubiquitin. Lysates were also immunoblotted with the indicated antibodies.

D, The mRNA levels of the indicated p53 target genes, including CDKN1A, SFN, PUMA, and BAX, were detected by real-time PCR in indicated cells stably expressing the control vector, Myc-COP1, or RFP-COP1 and normalized to GAPDH mRNA levels. Cells were also infected with the indicated Ad-β-gal (control) or Ad-14-3-3σ. The expression levels of the indicated p53 target genes were measured, and the data are presented as a heat map (Left). Statistic analysis for the heap map using Student’s t-test, p<0.05. The BDS2-3X-luc reporter containing a p53-responsive element was transfected with the indicated plasmids. Relative luciferase activity was shown (Right). Error bars represent 95% confidence intervals.