Repression of E2F transactivation by RIP140 and pocket proteins.

<p>(<b>A</b>) The human WT RIP900-luc reporter plasmid (25 ng) and E2F1+/− DP1 (25 ng each) were transiently transfected in MCF-7 cells with a dose response of RIP140 expression plasmids (0, 50, 100 or 200 ng). (<b>B</b>) ChIP experiments using immunoprecipitation (IP) of E2F1 and RIP140 on the E2Fa site of the human RIP140 promoter. The cyclin A2 promoter was used as a positive control. Data are expressed as percent of the PCR signal obtained with the amount of chromatin used for IP (input). Statistical analysis performed using the Student t test (***p<0.0001 and *p<0.05). (<b>C</b>) MCF-7 cells were transfected with human RIP900-luc or cyclin E-luc reporter plasmid (25 ng), with E2F1+/− DP1 (25 ng) and pocket proteins Rb or RIP140 expression vectors (200 ng or 250 ng) in 96 well plates in the indicated combinations. Results were expressed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035839#pone-0035839-g002" target="_blank">Figure 2D</a> (n = 4).