(A) Localization of putative HREs in the human PPARG gene. Gray and white rectangles represent noncoding and coding exons, respectively. Black rectangles represent putative HREs. Arrows mark localization of the PPARG isoform–specific promoters. (B) Relative enrichment of endogenous HNF1α at putative HREs on the PPARG and ALB genes in unsynchronized HUH7 cells. Data are means ± SEM, n = 3 independent experiments. *P < 0.05 vs. IgG; 2-tailed, unpaired Student’s t test. (C) The luciferase reporter activity normalized to β-gal activity in primary hepatocytes overexpressing HNF1α or GFP using HNF4A or PPARG promoter constructs. Data are presented as fold difference over empty vector–transfected AdGFP-infected condition. Data are means ± SEM, n = 3 independent hepatocyte cultures. *P < 0.05 vs. empty vector; 1-way ANOVA with Tukey’s multiple-comparisons test. (D) Immunoblot analysis of PPARγ protein after HNF1α overexpression in primary hepatocytes 24 hours after transduction with increasing doses of adenoviruses (0.5, 1, and 5 MOI). Immunoblot with anti-GAPDH antibody served as a loading control. (E) Luciferase activity of FGB-LUC, normalized to Renilla, measured in livers of 2-month-old male mice sacrificed 24 hours after hydrodynamic shock delivery. Data are means ± SEM, n = 3–7. *P < 0.05 vs. WT; ^#P < 0.05 vs. Pten LKO; 2-tailed, unpaired Student’s t test. (F and G) Immunoblot analysis of total protein extracts (F) and relative transcript levels (G) of HNF1α and HNF1α target genes in liver tissue of 4-month-old random-fed male mice. Densitometric analysis of actin-normalized signals is presented as a graph. Data are means ± SEM, n = 4–5. *P < 0.05 vs. WT; ^#P < 0.05 vs. Pten LKO mice; 2-tailed, unpaired Student’s t test.