Figure 5.

Foxa-2 regulates genes involved in glucose uptake, glycolysis, lipolysis, and energy dissipation. (a) Undifferentiated and differentiated 3T3-L1 cells containing the indicated expression vector were assayed for Hprt, Foxa-2, Foxc-2, Ir, Irs-2, Hsl, Lpl, Glut-4, M2Pk, Hk-2, Ucp-2, and Ucp-3 mRNA expression by RT-PCR. Fold regulation in undifferentiated, control 3T3-L1 cells (pcDNA3) versus Foxa-2–expressing cells: Ir: 3.3 ± 0.5*; Irs-2: 1.7 ± 0.3*; Lpl: 2.6 ± 0.4**; M2Pk: 3.3 ± 0.2***; Hk-2: 3.2 ± 0.3***; Ucp-2: 2.1 ± 0.7*; Ucp-3: 2.3 ± 0.4*. Expression levels of M2Pk were increased (3.4 ± 0.2**) in differentiated 3T3-L1–expressing Foxa-2 compared with control (pcDNA3). (b) Increased expression of metabolic genes in differentiated 3T3-L1 cells with enforced expression of Foxa-2 by adenoviral (Adeno) transduction. The 3T3-L1 cells were differentiated for 7 days and infected with control (no transgene) and recombinant adenovirus expressing Foxa-2. Cells were harvested after 48 hours, and gene expression was measured by RT-PCR. Experiments were carried out in triplicate. (c) Increased expression of metabolic enzymes in adipose tissue of ob/ob mice. Gene expression in fat tissue in ob/ob animals and wild-type littermates was measured by RT-PCR. Each lane indicates a different animal. Quantitative measurements of gene expression were obtained by densitometry, and ob/ob/WT indicates the ratio of adipocyte mRNA expression levels of the means of WT and ob/ob mice. The levels of significance of the comparison of WT versus ob/ob are shown on the right. *P = 0.05; **P = 0.01; ***P = 0.005.