Figure 3. Gck is a direct transcriptional target of LRH-1.

(A) Hepatic mRNA levels in Alb-Cre;Lrh1^fl/fl mice 5 weeks after in vivo transduction of the liver using AAV8-SHP virus (gray bars) in comparison with those in Lrh1^fl/fl mice (white bars) and Alb-Cre;Lrh1^fl/fl mice (black bars) (n = 4–5 per group). (B) Hepatic GCK protein expression in refed Lrh1^fl/fl and Alb-Cre;Lrh1^fl/fl mice. (C and D) Expression levels of LRH-1 and its targets in (C) wild-type primary hepatocytes and (D) Hepa 1.6 mouse hepatoma cells transduced with AdGFP (white bars) or AdLRH-1 (black bars) viruses (n = 3 per condition). (E) 2-deoxyglucose (2-DG) uptake and 2-deoxyglucose-6-phosphate (2-DG6P) production in Hepa 1.6 cells transduced with AdGFP (white bars) or AdLRH-1 (black bars) viruses (n = 6 per condition). (F) Schematic presentation of the 6 putative LRH-1 response elements in the mouse Gck promoter. (G) Assessment of LRH-1 recruitment to these sites, as depicted in F, determined by ChIP analysis using genomic DNA from livers of Lrh1^fl/fl and Alb-Cre;Lrh1^fl/fl mice. (H) Luciferase activities in HeLa cells transfected with empty luciferase reporter (pGL3; white bar) or long and short Gck promoter constructs (black bars). Data are expressed as fold induction in luciferase activity upon LRH-1 cotransfection. Data represent mean ± SEM. *P < 0.05 versus Lrh1^fl/fl, versus GFP, or versus empty reporter (pGL3); ^#P < 0.05 versus Alb-Cre;Lrh1^fl/fl.