Fig. 2

GREM2 inhibits adipogenesis in 3T3-L1 pre-adipocytes. A and B 3T3-L1 cells were cultured to confluence and differentiated using the standard induction cocktail with vehicle or GREM2 recombinant protein (50 ng/ml) for 9 days. The effect of vehicle or GREM2 protein on lipid accumulation was measured by Oil red O staining (A) and the bar graph represents relative lipid accumulation (B). E and F 3T3-L1-mock or 3T3-L1-Grem2 pre-adipocytes were cultured to confluence and differentiated using the standard induction cocktail for 9 days. The effect of mock or Grem2 overexpression in pre-adipocytes on lipid accumulation was measured by Oil red O staining (E) and the bar graph represents relative lipid accumulation (F). C and G On the 9th day of differentiation, proteins were isolated from adipocytes and the lysates were immunoblotted with the indicated antibodies. D and H The bar graphs represent relative band intensity. Two-sided t-test. ***, p < 0.001