Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2011 Aug 20.

Published in final edited form as: Circ Res. 2010 Jun 17;107(4):501–511. doi: 10.1161/CIRCRESAHA.110.222083

(A) HUVEC were infected with 25 PFU Ad-CMV-I| B(DN) or Ad-CMVGFP and stimulated with vehicle (PBS) or TNF⟨ (1 ng/ml) for 2 hours. NOR1 mRNA expression was analyzed and normalized to hTBP. Data are presented as mean ± SEM fold increase over Ad-CMV-GFP-infected cells treated with vehicle (*p < 0.05 vs. vehicle; #p < 0.05 vs. Ad-CMV-GFP). (B) Schematic structure of the human 4.0 kb and 1.7 kb NOR1 promoter constructs. (C) HAEC were transfected with a luciferase reporter construct driven by the indicated NOR1 promoter, stimulated with vehicle (PBS) or TNF⟨ (10 ng/ml), and analyzed for luciferase activities. Data are presented as mean ± SEM from three independently performed experiments (*p < 0.05 vs. vehicle; #p < 0.05 vs. pNOR1-1.7kb-WT). (D-E) HUVEC were stimulated with TNF⟨ (1 ng/ml) and harvested at the indicated time point for ChIP assays. Chromatin complexes were immunoprecipitated with an antibody against NF-| B p65 or species-matched IgG. PCR products were amplified using primers covering the −595 bp to −586 bp NF-| B binding site. (D) The autoradiograms are representative of three independently performed experiments. (E) Densitometric analysis of NF-| B p65 binding normalized to Input from three independently performed experiments.

(A) HUVEC were infected with 25 PFU Ad-CMV-I| B(DN) or Ad-CMVGFP and stimulated with vehicle (PBS) or TNF⟨ (1 ng/ml) for 2 hours. NOR1 mRNA expression was analyzed and normalized to hTBP. Data are presented as mean ± SEM fold increase over Ad-CMV-GFP-infected cells treated with vehicle (*p < 0.05 vs. vehicle; #p < 0.05 vs. Ad-CMV-GFP). (B) Schematic structure of the human 4.0 kb and 1.7 kb NOR1 promoter constructs. (C) HAEC were transfected with a luciferase reporter construct driven by the indicated NOR1 promoter, stimulated with vehicle (PBS) or TNF⟨ (10 ng/ml), and analyzed for luciferase activities. Data are presented as mean ± SEM from three independently performed experiments (*p < 0.05 vs. vehicle; #p < 0.05 vs. pNOR1-1.7kb-WT). (D-E) HUVEC were stimulated with TNF⟨ (1 ng/ml) and harvested at the indicated time point for ChIP assays. Chromatin complexes were immunoprecipitated with an antibody against NF-| B p65 or species-matched IgG. PCR products were amplified using primers covering the −595 bp to −586 bp NF-| B binding site. (D) The autoradiograms are representative of three independently performed experiments. (E) Densitometric analysis of NF-| B p65 binding normalized to Input from three independently performed experiments.