Fig. 5.

A schematic diagram (A) of the proximal promoter of the human NR0B2 gene with locations of putative insulin responsive elements (IRE) and Farnesoid X receptor response element (FXRE) relative to the transcription start site (TSS). DNA sequence alignments (B) among human, mouse, and rat Nr0b2 gene promoters in the four putative IREs with consensus sequences in uppercase. Luciferase reporter assays (C) using human NR0B2 gene promoter were performed in mouse H2.35 hepatocytes by transfection of the reporter constructs together with a single or combined constructs of GFP, FXR, Sirt1, Foxo1. *P < 0.05 for FXR, Sirt1, and Foxo1 vs. GFP overexpression; #P < 0.05 for cotransfection of two factors vs a single factor; **P < 0.05 for the triple transfection vs. Sirt1 cotransfection with Foxo1 or FXR. ChIP PCR analysis of Nr0b2 promoter association (D) was performed in mouse primary hepatocytes infected with GFP, Foxo1, or Sirt1 adenoviruses. DNA enrichment relative to the promoter of the internal control Ppia gene was quantified using real-time PCR. Values are presented as means ± SEM, n = 3. *P < 0.05 indicates a significant difference between enrichments by GFP and Sirt1or Foxo1.