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. Author manuscript; available in PMC: 2012 Sep 24.
Published in final edited form as: Free Radic Biol Med. 2011 Mar 26;50(12):1749–1759. doi: 10.1016/j.freeradbiomed.2011.03.022

FIGURE 9. Correlation between deletion of ARE sequences in NOX4 promoter and NOX4-Luciferarse reporter activity.

FIGURE 9

Seven deletions of ARE sequences in NOX4 promoter were carried out using Genscript’s (Piscataway, NJ) proprietary ClonEasy technology. The seven NOX4-Luciferase deleted reporter sequences were cloned into pGL3 vector, and sequences were confirmed with the gene sequencing facility at the University of Chicago. HLMVECs (~80% confluence) were transfected with these 7 ARE deleted plasmids (~1μg with Fugene HD transfection reagent) for 24 h as described in “Materials and Methods”. After 24 h, the cells were exposed to normoxia or hyperoxia for 3 h and cell lysates were analyzed for luciferase activity using Promega Dual Luciferase kit. Values are mean ± S.E.M for three independent experiments in triplicate. *, significantly different from normoxia (p<0.05); **, significantly different from NOX4-Luciferase (-438-458, -619-636 and -665-677) transfected cells exposed to hyperoxia (p<0.01).