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. Author manuscript; available in PMC: 2010 Apr 28.
Published in final edited form as: Circulation. 2009 Apr 13;119(16):2161–2169. doi: 10.1161/CIRCULATIONAHA.108.815803

Figure 5. Functional analysis of the PPRE located in the β2-AdR promoter.

Figure 5

(A) PPARγ binds to the putative PPARγ element in the mouse β2-AdR promoter. Cells were transfected with pcDNA3.1 carrying Flag-tagged PPARγ for 48 hours. ChIP assay of the mouse β2-AdR promoter was performed using anti-Flag antibody. DNA from immunoprecipitated β2-AdR chromatin and input was subjected to PCR analysis using two pairs of primers covering regions containing the putative PPRE as well as at -15 kb (control) in the mouse β2-AdR promoter. The experiments were repeated at least three times. (B) Dual-luciferase reporter assays were performed transfecting VSMC with a β2-AdR wild-type promoter (β2-WT) versus a PPRE mutated β2-AdR promoter (β2-mu) generated by site-directed mutagenesis. VSMC were co-transfected with pcDNA3.1 and Flag-PPARγ and treated with GW7845 overnight. β2-AdR promoter activity was reduced upon GW7845 treatment but not in the β2-mu promoter. Expression of Flag-PPARγ reduced β2-WT promoter activity but not that of β2-mu. Data are shown as mean±S.E.M, (n=6).