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. Author manuscript; available in PMC: 2013 Mar 16.
Published in final edited form as: Circ Res. 2012 Feb 21;110(6):808–817. doi: 10.1161/CIRCRESAHA.111.259325

Figure 4. ATF6 binds to sEH promoter and activates promoter activity.

Figure 4

(A) Homology analysis of ATF6 binding site on the promoter of sEH among of human, rat and mouse by UCSC Genome Browser. Plasmids of sEH-1000-Luc or ATF6 site mutation (Mu283) were transfected into EA.hy926 cells, and then treated with Hcy, with or without AEBSF in (B), or infected with Ad-GFP or Ad-ATF6(N) for 24 hr in (C). Serial deletion constructs of sEH-Luc were transfected into EA.hy926 cells and treated with Hcy, with or without AEBSF for 24 hr (D). The β-gal plasmid was co-transfected as a transfection control. Promoter activities were measured by luciferase activity, which was normalized to that of β-galactosidase. Data are mean±SD of relative luciferase activities from 4 independent experiments. (E) ChIP assay involved use of anti-ATF6 antibody for immunoprecipitation in treated HUVECs; normal rabbit IgG was a control. Semi-quantitative PCR involved sEH promoter-specific primers to detect the binding of ATF6 to the sEH promoter region. Data are mean±SD of the percentage of input control from 3 independent experiments. *P<0.05, **P<0.01 vs. Ctrl; #P<0.05 vs. Hcy or Ad-ATF6.