Figure 6.

Foxa-2 is a transcriptional activator of Pref-1, Ucp-2, Lpl, and Hk-2 gene promoters in vitro. (a) Foxa-2 transactivates the promoters of Ucp-2, Lpl, Pref-1, and Hk-2 in 3T3-L1 cells. The 3T3-L1 cells were transfected with the indicated expression vectors, pCMV–β-gal, and the luciferase reporter genes. Luciferase activity was normalized to β-gal activity. Each value represents mean of nine independent experiments ± SD. *P < 0.01; **P < 0.001. (b) Foxa-2 from ob/ob fat nuclear extract binds to sites in the Pref-1, Lpl, and Ucp-2 promoters. ³²P-labeled probes corresponding to putative Foxa-2–binding sites in the promoters of Pref-1, Lpl, and Ucp-2 were incubated with nuclear extracts from ob/ob fat in the presence of either unlabeled probe, anti–Foxa-1, or anti–Foxa-2 Ab (21). Protein/DNA complexes were separated on a 4% acrylamide gel and visualized by autoradiography. The radioactive probes (bottom of the gel) are not shown.