BMP-6 represses binding of endogenous δEF1 to the E-cadherin proximal promoter in MDA-MB-231 breast cancer cells. (a) Association of δEF1 with the E-cadherin promoter region was examined in vivo by CHIP analysis in MDA-MB-231 cells, using an unrelated anti-FLAG antibody or an anti-ZEB antibody directed against the N-terminal epitopes of δEF1. The amplified human E-cadherin promoter fragment is shown (-175/+21). (b) For quantitative CHIP assay, MDA-MB-231 cells were treated with or without 200 ng/ml rhBMP-6. Cell lysates were collected after 72 h. The IP was performed using anti-ZEB antibody (10 μg) with anti-FLAG antibody (10 μg) as a negative control. DNA fragments containing the E-cadherin promoter region (-175/+21) were amplified by quantitative PCR from anti-ZEB and anti-FLAG immunoprecipitated samples. Data represent three independent experiments. * indicates p < 0.05 in unpaired t-test when compared with un-treated group.