ERalpha CD domain interacts with Pin1.

<p>A) Schematic representation of ERalpha domains, showing the predicted prolyl isomerase sites (Ser/Thr-Pro). B) ERalpha cDNA was split into three different domains (AB, CD, EF) and tagged with histidine. GST or GST-Pin1 pull-down assay was performed in 293FT cells and analyzed by Western blot with anti-6×His antibody. C) Site-directed mutagenesis was performed to replace Serine 294 with Alanine on the ERalphaCD domain. Plasmids with wild-type (w) or mutant (m) CD His-tagged domain were transfected in 293FT cells and pulled down with GST-Pin1 protein. Anti-His antibody was used in Western blots to detect the interaction.