Figure 9. E2F1–E2F4 bind to the ASK-1 promoter in vivo.

Formaldehyde cross-linked chromatin from He La (0) cells were subjected to ChIP experiments. Immunoprecipitations of E2F containing complexes were performed using anti-E2F1 (α-E2F1), anti-E2F2 (α-E2F2), anti-E2F3 (α-E2F3) and anti-E2F4 (α-E2F4) antibodies. Anti-ICAM1 polyclonal antibody (α-ICAM1) was used as negative control and anti-(RNA pol II) (α-RNA pol II) as a positive control. After isolation of bound DNA, PCR was performed for a 297 bp region of the endogenous ASK-1 promoter. As positive controls, PCRs were also performed using E2F1-promoter-specific primers and CCNA2-promoter-specific primers, two well-documented E2F target genes, and ASK-1-intron 5-specific primers as a negative control. Input indicates PCR performed on DNA (diluted 1/300) without any immunoprecipitation. Values on the right indicate the number of PCR cycles performed. prom, promoter.