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. Author manuscript; available in PMC: 2008 May 12.
Published in final edited form as: Nat Cell Biol. 2008 Jan 20;10(2):138–148. doi: 10.1038/ncb1676

Figure 3.

Figure 3

MDM2 is required for ERk-mediated FOXO3a degradation. (a) MCF-7 cell lysates were subjected to immunoprecipitation with an anti-MDM2 antibody and immunoblotting with an anti-FOXO3a antibody. (b) Lysates of 293T cells transfected with MDM2 at different dosages and subjected to immunoblotting. (c) Lysates of cotransfected cells treated with the indicated vectors in the presence of MG-132 (20 μM, 4 h) were subjected to immunoprecipitation and immunoblotting with an anti-FOXO3a antibody. (d) Lysates of p53−/− or p53−/−MDM−/− MEFs were subjected to immunoblotting using different antibodies, as indicated. (e) MEFs were treated with different inhibitors and analysed by immunoblotting. (f, g) MEFs were treated with or without CHX, harvested at the indicated times and subjected to immunoblotting. (h) MEFs were treated with methionine and cysteine-free medium overnight, pulsed with 35S-Met-Cys for 30 min, and chased for the indicated times. Cell lysates were immunoprecipitated with an anti-FOXO3a antibody and subjected to SDS–PAGE, gels were fixed, dried,and subjected to autoradiography (with an intensifying screen). An uncropped image of the blot in e is shown in the Supplementary Information, Fig. S5.