Figure 7.

Effect of chimeric Sox2/11 overexpression on Sox2 and Nanog promoter activity. (A) Schematics of chimeric Sox constructs. Sox2-2-11 contains the Sox2 DBD coupled with the Sox11 TAD whereas Sox11-11-2 contains the Sox11 DBD coupled with the Sox2 TAD. (B) Effect of chimeric Sox overexpression on Sox2 promoter activity. (C) Effect of chimeric Sox overexpression on Nanog promoter activity. In both (B) and (C), a schematic of the promoter/reporter construct is 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. F9 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, 3 µg of either the Sox2, Sox2-2-11 or Sox11-11-2 expression vector was co-transfected. Reporter activity was normalized to that of β-galactosidase and the activity of each individual promoter/reporter construct (with no 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 twice and similar results were obtained.