Pin1 associates with ERalpha both <i>in vitro</i> and <i>in vivo</i>.

<p>A) Serum-starved cells were treated with estradiol (E2) or ethanol for 45′. Cells were harvested and pellets immunoprecipitated with anti-Pin1, and analyzed by Western blot with anti-ERalpha antibody. Pin1 kd cells or IgG were used as a negative control. B) MCF7 cells were transfected with shRNA scramble (SCR) or shRNA Pin1 (kd1 and kd2), and Pin1 protein level was detected by immunoblotting. GAPDH was used as the load control. Quantification represents the ratio between the Pin1 and GAPDH proteins and normalized to scrambled cells. C) GST, GST-Pin1, GST-WW or GST-PPIase fusion proteins were incubated with ERalpha-positive MCF7 cell lysates and the bound proteins were analyzed by immunoblotting with an ERalpha antibody.