Figure 12

Importance of the AP-1 element (-99 to -93) for the Tkdp-1 .140 bp promoter activity in JEG-3 cells. A) Effects of the AP-1 element mutation on the basal, activated PKA, Ets-2 and Ets-2+activated PKA-dependent activation of the 140 bp Tkdp-1 promoter-Luc reporter construct in JEG-3 cells. Luc activities are expressed relative to that of the basal activity of the 140 bp promoter and data are shown as mean ± SEM (n = 3). Following values are significantly different at P < 0.05: 140+PKA vs. AP-1mut140+PKA and 140+Ets-2+PKA vs. AP-1mut140+Ets-2+PKA. B) and C) Pull-down assays with biotinylated AP-1 oligonucleotide (-99 to -93 in the tkdp-1 promoter) and JEG-3 cell extracts(CE). Following pull-down a ssays, WB analysis was performed with B) C/EBP-β IgG and C) pan-Jun IgG. Lane 1: JEG-3 CE (positive control for WB); lane 2: biotinylated AP-1 oligonucleotide; lane 3: JEG-3 CE+biotinylated AP-1 oligonucleotide. The molecular weight standards are shown on the left-hand side.