Figure 3

ZEB1 associates with estrogen receptor in MCF-7 cells. MCF-7 cells were treated with PBS/0.1% ethanol control vehicle, 1 nM estrogen, 10 nM tamoxifen or with estrogen in combination with tamoxifen for 24 h. ZEB1 was immunoprecipitated from cell lysates using anti-ZEB1 antibodies, as described in “Methods”. Immunoprecipitated proteins were separated by SDS-PAGE and transferred to PVDF membranes. Presence of ER α in immunoprecipitated samples was determined by Western immunoblotting using anti-ER α antibodies, as described in “Methods”. Imaging of membranes was performed with a Versadoc imager and Multi-analyst software.