Figure 7.

Human Cited2 transcription levels also respond to changes in E2F1/4 levels. Mutations of both E2F binding sites significantly decrease the effect of E2F1 levels over Cited2 transcription. A, features of the human Cited2 locus. Two putative E2Fs binding sites are identified, and mutations were introduced to generate two individual reporter constructs (hRSmut1–2). B, HEK293T cells were co-transfected with Renilla luciferase as control and firefly luciferase under the control of the specified (native, hRS-Luc, mutated hRSmut1-Luc, or mutated hRSmut2-Luc) human Cited2 genomic regulatory sequence. Cited2 transcription levels were then measured as luciferase activity after ectopic E2F1 expression. Two-way ANOVA/Tukey's were used to analyze the data. Only some of the Tukey's tests are shown, although all comparisons between GFP and E2F1 groups revealed significant differences. hCited2, human Cited2; RS, regulatory sequence; mut, mutant; Luc, luciferase.