FIGURE 3.

E2F1 binds to the Bim promoter at multiple sites. A and B, putative E2F1-binding sites on the Bim promoter were determined by in silico analysis using KTSSCGC consensus sequence. C–F, ChIP analysis determines multiple E2F1-binding sites on the Bim promoter. Briefly, cancer cells were subjected to chromatin cross-linking followed by immunoprecipitation with anti-E2F1, anti-Sp1, anti-FOXO3a, and RbIgG. Bim promoter fragments were amplified by using PCR primers as described in Table 1. Positive controls (input DNA) or negative controls (H₂O) were used in the amplification of the Bim promoter. Crossed sites in A (i.e. 1–5 and 9 predicted sites) indicate that these sites do not show binding with E2F1 based on ChIP analysis.