Chromatin immunopreciptation analysis.

<p>E2F1 binds to the CD2AP promoter in vivo. HEK293 cells were transfected with pcDNA3 or pcDNA3-E2F1 expression vector. 24 h post-transfection, cells were harvested and subjected to ChIP assay. ChIP products were amplified by PCR reaction. Cross-linked chromatin samples from pcDNA3-transfected HEK293 cells, pcDNA3-E2F1-transfected HEK293 cells were respectively, immunoprecipitated with anti-E2F1, and normal rabbit IgG (anti-IgG). Purified DNA was analyzed by PCR using primers as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042774#s2" target="_blank">Materials and methods</a>. A sample representing 10% of the chromatin used as a starting material for the immunoprecipitation is also shown (input). The amplified PCR fragments were analyzed on 2.0% agarose gel. Each experiment was performed twice with similar results, and one of them is shown.