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. 2013 Apr 5;41(10):5210–5222. doi: 10.1093/nar/gkt223

Figure 6.

Figure 6.

Arsenite exposure enhances S7–MDM2 interaction, which subsequently abrogates GADD45α ubiquitination. (A) HepG2 cells were either transfected with HA–MDM2 or in combination with GFP–S7. Then the cells were left untreated or exposed to arsenite (20 µM) for 8 h. Cell lysate was immunoprecipitated with anti-GFP antibody, and the immunoprecipitants were probed with anti-GFP and anti-MDM2 antibodies. (B) HepG2 cells were left untreated or pre-treated with MG132 (10 µM) for 6 h followed by exposure to arsenite (20 µM) for 8 h. Cell lysate was immunoprecipitated with anti-S7 antibody or the mouse IgG, and then the immunoprecipitants were probed with anti-S7 and anti-MDM2 antibodies. (C) HepG2 cells were transfected with Myc–Ub expression plasmid with or without combination of FLAG–GADD45α or HA–MDM2 constructs. Then the transfected cells were left untreated or exposed to arsenite (20 µM) for the time indicated. Cell lysate was immunoprecipitated with anti-FLAG antibody, and then the ubiquitination of GADD45α was detected with anti-GADD45α antibody.