WTX and TRIM28 regulate adipogenic differentiation.
A, adipogenic differentiation of preadipocyte 3T3-L1 cells following knockdown of WTX or TRIM28. Lipid formation was visualized with oil red-O staining after culture under adipogenic differentiation conditions for 12 days. Where indicated, PNU-74654 (PNU) was added to 100 μm final concentration. Magnified images of each well in the upper panel are shown in the lower panel with hematoxylin counter staining. B, expression of adipogenic marker genes Pparγ and C/ebpα in 3T3-L1 cells cultured under adipogenic condition for 8 days. Control cultures were maintained under non-induced (NI) condition. **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001 compared with control (shGFP). C, chromatin binding of TRIM28 on Pparγ, C/ebpα, and Mest promoters in 3T3-L1 cells cultured under non-induced (NI) or adipogenic (Adipo) conditions determined by ChIP-quantitative PCR following knockdown of WTX or control GFP. ns, p > 0.05; *, p ≤ 0.05 compared with control (shGFP). Error bars represent S.E. IP, immunoprecipitation.