Fig. 4.

Inhibition of GSK3β suppresses adipogenesis by induction of Snail expression in 3T3-L1 cells. a Immunoblot analysis shows that Wnt10b increases the expression of phosphorylated GSK3β and Snail in preadipocytes during adipogenic differentiation. Transfected 3T3-L1 cells were cultured for the indicated times before and after initiating the treatment with adipogenic hormonal cocktail. Actin was used as a loading control. b Immunoblot analysis of differentiated adipocytes treated with the indicated reagents on day 8. The expression levels of phosphorylated GSK3β and Snail are significantly increased in LiCl-treated cells. Cells with no reagent and NaCl-treated cells were used as controls. Actin was used as a loading control. c On day 8 after treatment with adipogenic cocktail, control and cells treated with LiCl (40 and 70 mM) or NaCl (40 mM) were stained with Oil Red O. Cells with no reagent and NaCl-treated cells were used as controls. To assess the role of Snail in the GSK3β-mediated suppression of adipogenesis, 3T3-L1 preadipocytes transfected with Snail siRNA or scrambled control siRNA were treated with LiCl (40 mM). On day 8, after the treatment with adipogenic cocktails, Snail siRNA-transfected cells differentiated into adipocytes, confirmed by Oil Red O staining (d) and immunoblots (e). The figures are representative of three independent experiments. f Densitometric graph of the optical density-based data of the immunoblots. *p < 0.001 vs. compared to the cells with control siRNA treatment (control). Data are presented as the mean ± SD of three independent experiments