Way 200070

Parental and ERβ1-expressing MCF7 cells [12 (link)] and doxycycline-inducible Hs578T-ERβ1 cells [8 (link)] were cultured as previously described. Doxycycline-inducible ERβ1-expressing MDA-MB-231 cell lines were established using the T-REx™ System (Invitrogen) as previously described [9 (link)] and were maintained in DMEM/F12 medium supplemented with 10% FBS, 1% AA, 5 mg/L blasticidin S and 500 mg/L zeocin. Charcoal-stripped fetal bovine serum (CS-FBS) was purchased from Gemini Bio-Products (West Sacramento, CA). 17β-estradiol (E2), (Z)-tamoxifen, (Z)-4-hydroxy-tamoxifen and doxycycline (Dox) were purchased from Sigma-Aldrich (St. Louis, MO). (Z)-endoxifen was synthesized by the National Cancer Institute (Bethesda, MD). The ERβ-specific agonists; DPN, WAY200070, FERb 033 and Liquiritigenin, as well as the pure ER antagonist ICI 182,780, were purchased from Tocris Bioscience (Bristol, United Kingdom).

The human ovarian cancer cell line OVCAR-3 was obtained from American Type Culture Collection (ATCC #HTB-161, Manassas, USA), and OAW-42 ovarian cancer cells were obtained from Sigma Aldrich (#85073102, St. Louis, USA). The cells were maintained in phenol red-free DMEM culture medium that was obtained from Invitrogen (Karlsruhe, Germany) containing FCS that was purchased from PAA (Pasching, Austria). RNeasy Mini Kit was obtained from Qiagen (Hilden, Germany). Transfectin reagent was obtained from BioRad (Hercules, USA). OptiMEM medium were purchased at Invitrogen (Karlsruhe, Germany). ESR2 and control siRNAs were from Ambion (Life Technologies, USA). Serum Replacement 2 (SR2) cell culture supplement and 17-β estradiol were from Sigma-Aldrich (Deisenhofen, Germany). ERβ agonists ERB-041 and WAY-200070 were from Tocris (Bristol, UK). 5α-androstane-3β, 17β-diol (3β-Adiol) was from Sigma (Deisenhofen, Germany) and Liquiritigenin from Extrasynthese (Lyon, France).

Phenol red-free DMEM culture medium was obtained from Invitrogen (Karlsruhe, Germany), FCS was purchased from PAA (Pasching, Austria). MDA-MB-231 and HS578T breast cancer cells were obtained from American Type Culture Collection (Manassas, USA). RNeasy Mini Kit was obtained from Qiagen (Hilden, Germany). Transfectin reagent was obtained from BioRad (Hercules, USA). OptiMEM medium were purchased at Invitrogen (Karlsruhe, Germany). ESR2 and control siRNAs were from Ambion (Life Technologies, USA). Serum Replacement 2 (SR2) cell culture supplement was from Sigma-Aldrich (Deisenhofen, Germany). ERβ agonists ERB-041 and WAY-200070 were from Tocris (Bristol, UK). 5α-androstane-3β, 17β-diol (3β-Adiol) was from Sigma (Deisenhofen, Germany) and Liquiritigenin from Extrasynthese (Lyon, France).