Figure 2.

Foxa-2 is an inhibitor of adipocyte differentiation. (a) Inhibition of adipocyte differentiation in 3T3-L1 cells. Cells were transfected with vector pcDNA3 (control) or expression vectors containing cDNAs of Foxa-1, Foxa-2, and Pref-1. Pools of stable transfectants were induced with differentiation medium (not containing insulin). At day 8 after induction, cells were either stained for lipid accumulation using oil red O or mRNA, and total protein extracts were prepared. (b) Measurements of gene expression profiles using RT-PCR. Hprt expression was used as a loading control indicating that each sample contained similar amounts of mRNA. No products were amplified in the absence of reverse transcriptase. (c) Western blot analysis of cell extracts from undifferentiated and differentiated 3T3-L1 cell lines. Total protein was separated by SDS-PAGE and analyzed by immunoblotting for Foxa-2 and aP2 expression. TATA-binding protein (Tbp) expression was measured as a loading control. (d) Differentiation of primary preadipocytes of wild-type, mutant Foxa-2 (Foxa-2^+/–), and ob/ob cells. (e) Gene expression of Foxa-2, Pref-1, and aP2 in differentiated primary preadipocytes of Foxa-2^+/–, wild-type, and ob/ob mice.