Mibolerone mib

11KT, 11KDHT and mibolerone (Mib) were purchased from Steraloids. T and DHT were purchased from Sigma-Aldrich, while [H³]-Mib (76.8 Ci/mmol) was purchased from Perkin Elmer. All steroids were dissolved in absolute ethanol and added to the culturing medium at a final concentration of no more than 0.1% ethanol. Cortisol-9, 11, 12, 12-d4 (cortisol-d9), testosterone-1, 2-d2 (T-d2), progesterone-2, 2, 4, 6, 6, 17α, 21, 21, 21-d9 (P4-d9) and 4-pregnen-17α-ol-3,20-dione-2, 2, 4, 6, 6, 21, 21, 21-d8 (17OHP4-d8) were purchased from Cambridge Isotope Laboratories. Gestodene (GES) and drospirenone (DRSP) were purchased from Sigma-Aldrich.

Expression plasmids, including pcDNA3.1(+)-CXCL13, pcDNA3.1(+)-AR, pcDNA3.1(+)-AR-N, pcDNA3.1(+)-FOXO1 and pcDNA3.1(+)-HDAC1, were constructed by and stored in our lab. Series of dual-luciferase test plasmids of AR binding elements in CXCL13 enhancer (including wild-type and mutants) were constructed in pGL3-promoter vector (purchased from Promega, WI, USA). For CXCL13 stable-expression cell line, the plasmid pcDNA3.1(+)/CXCL13 was transfected into CWR22Rv1 cells. After 48 hr, cells were cultured in medium containing G418 for the selection of stable clones.
Antibodies against AR (N-20), AR(441), FOXO1, HDAC1, β-actin, ETS-1 and Cyclin B1 were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Snail was assessed with an antibody from Cell Signaling Technology, and CXCL13 with an antibody from Sigma-Aldrich (Sigma-Aldrich, Munich, Germany). The appropriate secondary antibodies were all purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). All siRNAs (including CXCL13 specific siRNA, AR specific siRNA and control siRNA) were purchased from Santa Cruz Biotechnology Inc. Mibolerone (Mib) was purchased from Steraloids Inc. (Newport, RI, USA). Other chemicals were all purchased from Sigma-Aldrich. The molecular weight (protein size) of same proteins was labeled when it first appeared in figures.