Figure 2. Repression of PKA–CRE-mediated activity.

The −259 bp G6Pase promoter–luciferase reporter plasmid and its mutant constructs were co-transfected with pCR3/hPXR and/or pCR3/hPKA, and the reporter activity was measured as described in the Experimental section. (A) The −259 bp wild-type promoter, (B) the −259 bp_irs/hnf4 promoter bearing the double mutations of IRS and HNF4 sites, (C) the −259 bp_{irs/cres/hnf4} promoter having the triple mutations at the IRS, two CRE and HNF4 sites. pRL-CMV was always co-transfected for the control. Total amount of DNA was adjusted by adding pcDNA3-V5-His as an empty vector control. Relative fold activities were calculated by taking the activity of the cells that were transfected by the reporter plasmid only in the presence of DMSO as 1. Results are means±S.D. for three experiments.