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. Author manuscript; available in PMC: 2013 Sep 4.
Published in final edited form as: Arch Biochem Biophys. 2011 Apr 21;511(0):31–39. doi: 10.1016/j.abb.2011.04.009

Figure 2. Serum activates the proximal promoter of the human nCDase gene.

Figure 2

A) Deletion analysis of the 5′-untranslated region of the human nDase gene verified the major positive regulatory elements of the human nCDase gene reside within a 200 bp minimal essential promoter region. Luciferase expression vectors containing different regions of the nCDase promoter were cotransfected into HEK 293 cells with a plasmid (phRL-null) containing the renilla luciferase gene. Following 24 h incubation with DMEM containing FBS, firefly luciferase (fLuc) and renilla luciferase (rLuc) activities were measured. The fLuc/rLuc ratios were determined, and means for a minimum of three transfections were calculated. Values are expressed as mean ± S.E.M-fold increase relative to the luminescence ratio observed in pGL3-Basic transfected cells (relative value = 1). B) Luciferase activity of the proximal promoter is increased through serum treatment. HEK 293 cells cotransfected overnight with PGL-200, the 200 bp promoter/reporter construct, were serum starved for 24 hours and then treated with serum or basal media for the indicated time points. Values are expressed as mean ± S.E.M-fold increase in luminescence relative to serum starved PGL-200 transfected cells (relative value = 1). C) The minimal essential promoter region of the human nCDase gene contains a major potential binding site for the serum regulated transcription factor, AP-1. Nucleotide sequence of the region from −200 to −1 was analyzed for consensus transcription factor-binding elements using TESS analysis. Putative cis-elements are underlined or over scored (over-lined) in the case of overlapping sites.