Figure 8.

Effect of Sox2 overexpression on Sox2:Oct-3/4 target genes in P19 EC cells and D3 ES cells. (A) Effects of Sox2 overexpression on Sox2, FGF-4, Nanog, UTF1 and Oct-3/4 promoter/reporter gene constructs in P19 EC cells. (B) Effects of Sox2 overexpression on Sox2, FGF-4, Nanog and Oct-3/4 promoter/reporter gene constructs in D3 ES cells. Schematics of each of the promoter/reporter constructs are shown. The gray boxes indicate critical enhancers, the basal promoter is represented by a black box adjacent to the CAT reporter gene, and the relative HMG/POU cassette is indicated with an asterisk. In (A), P19 EC cells were transfected with 12 µg of the indicated promoter/reporter gene construct plus 1 µg of the CMV-β-gal expression vector. Where indicated, the cells were also transfected with 1 or 3 µg of the Sox2 expression vector. In (B), D3 ES cells were transfected with 4.5 µg of the indicated promoter/reporter gene construct plus 1 µg of the CMV-β-gal expression vector. Where indicated, the cells were also transfected with 0.3 or 1 µg of the Sox2 expression vector. In both (A) and (B), reporter activity was normalized to that of β-galactosidase and the activity of each individual promoter/reporter construct (with no Sox2 overexpression) was set to 1. Overexpression values are shown relative to each particular gene construct. The data shown represents the mean and standard deviation of duplicate samples from a representative experiment. Each experiment was repeated and similar results were obtained.