Figure 5.

Overexpression of Dnmt3a rescues the impairment of adipogenesis induced by AP2α knockdown. (a) 3T3-L1 preadipocytes at ci0 h were transiently transfected with either scramble siRNA (ctrl) or AP2α-specific siRNAs (si-1 and si-2), and then induced with hormone cocktail at ci48 h. On day 8, the cells were stained with Oil-Red-O (left panel), and then extracted with isopropanol and measured at OD_510 nm. The data were presented as means±S.D. (n=3) and shown on the right panel. Statistical significance is indicated: *P<0.01. (b) Western blotting analysis of adipogenic factors on the cells induced by MDI for 8 days. (c) For rescue study, 3T3-L1 cells at ci0 h were transiently co-transfected with three pairs of plasmids (scramble siRNA (ctrl) and GFP plasmid, AP2α-specific siRNA (si-1) and GFP plasmid, AP2α-specific siRNA (si-1) and Dnmt3a plasmid), respectively. The cells were then induced with hormone cocktail after 48 h transfection. On day 8 of MDI induction, the cells were stained with Oil-Red-O (top panel) and extracted with isopropanol and measured at OD_510 nm (bottom panel). The data were presented as means±S.D. (n=3). (d) Western blotting analysis of adipogenic factors on the cells induced by MDI for 8 days. (e) The expressions of Dnmt3a and AP2α were analyzed with western blotting. Cyclin-dependent kinase 4 served as the loading control