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. 2011 Dec;31(24):4951–4963. doi: 10.1128/MCB.05553-11

Fig. 9.

Fig. 9.

p53 misfolding function of MDM2 is regulated by the ATM sites. (a) H1299 cells were transfected with p53 and MDM2 for 24 h and treated with 30 μM MG132 for 5 h to inhibit p53 degradation. Cells were also treated with 7 Gy of IR, as indicated, 4 h before harvesting. Cell lysates were immunoprecipitated with wild-type conformation-specific (Pab1620) or mutant conformation-specific (Pab240) antibodies. The precipitated p53 was detected by Western blotting with FL393 antibody. Whole-cell extracts (WCE) were analyzed for protein levels. (b) SJSA (amplified MDM2) and U2OS cells were treated with 30 μM MG132 for 5 h to inhibit p53 degradation. Cells were also treated with 7 Gy of IR as indicated 4 h before harvest or with 5 nM actinomycin D for 16 h. Cell lysates were immunoprecipitated with Pab1620 or Pab240. The precipitated p53 was detected by Western blotting with FL393. WCE were analyzed for protein levels.