On target pooled sirnas

On-Target pooled siRNAs, miRNA mimics/inhibitors, and transfection reagents were obtained from Thermo-Fisher Scientific (Pittsburgh, PA, USA); pmiR-GLO luciferase vector and Dual Luciferase Reporter assay were obtained from Promega (Madison, WI, USA). shRNA vectors were from the University of Colorado Functional Genomics Facility (Boulder, CO, USA). Primary antibodies used included: Stat5 (SC-835, SC-836; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), CK5 (mouse NCL-L-CK5 and rabbit 2290-1; Vector Laboratories/Epitomics, Burlingame, CA, USA), PR (Dako, Carpinteria, CA, USA), α-tubulin and β-actin (Sigma-Aldrich, St. Louis, MO, USA). For western blot, secondary antibodies included: IRDye 800CW Goat-Anti-Mouse IgG (926-32210) and IRDye 680LT Goat-Anti-Rabbit IgG (926-68021)(Li-Cor Biosciences, Lincoln, NE, USA). For Western blot imaging, the Odyssey Infrared Imaging System (Li-Cor Biosciences). For immunocytochemistry, secondary antibodies: AlexaFluor488 goat anti-mouse (A11029) and goat anti-rabbit (A11008), AlexaFluor594 goat anti-mouse (A11032) and goat anti-rabbit (A11037) (Invitrogen/Life Technologies, Grand Island, NY, USA). Hormones (17β-estradiol, progesterone, and MPA) were purchased from Sigma. Pimozide (573110) was purchased from EMD Millipore (Billerica, MA, USA).