Fig. 3.

Wnt10b blocks adipogenesis by induction of Snail expression in 3T3-L1 cells. a On day 8 after treatment with adipogenic cocktail, the control cells and cells transfected with plasmids harboring Wnt10b were stained with Oil Red O. These data are representative of three independent experiments. b Immunoblot analysis shows that Wnt10b increases the expression of Snail in preadipocytes and during adipogenic differentiation. The 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. c The graph shows densitometric analysis of the optical density-based data of the immunoblots shown in Fig. 4b. It can be seen that Wnt10b increases the expression of Snail in 3T3-L1 cells, regardless of differentiation. *p < 0.001 vs. compared to the cells without Wnt10b treatment (control). Data are presented as the mean ± SD of three independent experiments. d To assess the role of Snail in the Wnt-mediated suppression of adipogenesis, 3T3-L1 preadipocytes transfected with Wnt10b-expressing plasmids were treated with Snail siRNA or scrambled control siRNA. On day 8, after the treatment with adipogenic cocktails, Snail siRNA-transfected cells show full differentiation into adipocytes. e Immunoblot analysis of Snail, C/EBPα, PPARγ and adiponectin expression in 3T3-L1 cells on day 8 after treated with either Snail or control siRNA. The data 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