FIGURE 7.

Effects of rosiglitazone on the ubiquitination of anti-adipogenic proteins. C3H10T1/2 cells were treated with rosiglitazone (R) (10 μm) in the presence or absence of PHD inhibitors for 10 days. Whole cell extracts were immunoprecipitated (IP) with anti-polyubiquitination antibody, and this was followed by Western blotting using anti-GATA-3, anti-KLF-2, or anti-TAZ (A and B). As a control, immunoprecipitation was carried out using mouse IgG antibody (C). Separately, whole cell extracts were analyzed by Western blotting for these anti-adipogenic proteins. D, DMOG; E, EDHB; Ub, ubiquitin; C, control. MG-132 (5 μm) was added in differentiation medium at the end of the 10-day differentiation protocol, incubated for either 6 or 24 h, and then whole cell extracts were analyzed by Western blotting (C). Experiments were carried out twice, and representative results are shown.