Crl 2505

Human prostate cancer cell line PC3 (CRL1435; ATCC), 22RV1 (CRL2505; ATCC), monocytic leukaemia cell line THP‐1 (TIB‐202; ATCC) and mouse fibroblast cell L929 (CCL1; ATCC) were provided by Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai, China. PC3 and 22RV1 were maintained in RPMI 1640 supplemented with 5% (v/v) foetal bovine serum (FBS, 10099141; Gibco). THP‐1 was grown in RPMI 1640 medium supplemented with 5% (v/v) FBS, 200 μmol/L glutamine (Thermo Fisher Scientific) and 0.2 μmol/L β‐mercaptoethanol (Sigma). L929 cells were maintained in Dulbecco's modification of Eagle's medium (DMEM) supplemented with 5% (v/v) FBS.
Bone marrow‐derived macrophage (BMDM) was isolated from mouse tibia according to Spring Harbor Protocols.³³ In brief, femur and tibia bones are collected from 6 to 8 weeks male C57BL6/J mice and bone marrow cells were flushed out using Hank's buffer. After lysis of red blood cells. BMDM cells are cultured in BMDM growth medium (70% DMEM complete medium, 30% L929 cell supernatant).

All cell lines were cultured at 37 °C with 5% CO₂. The medium used for VCaP (CRL-2876, ATCC), 22RV1 (CRL-2505, ATCC), V16A (established by Dr. Amina Zoubeidi’s laboratory), and HEK293FT (R70007, Thermo Fisher Scientific) cell culture was supplemented with 10% FBS (GIBCO, 10437-028), 1% streptomycin and 1% penicillin. For androgen stimulation treatment, cells were grown to 50%~60% confluence in a medium containing 5% charcoal-dextran stripped FBS (CDS) for 48 h and then treated by 10 nM DHT for 2 or 24 h. All cell lines used in this study were tested negative for mycoplasma contamination. We utilized ATCC services following extended passages to authenticate by utilizing Short Tanden Repeat (STR) profiling.

CWR22Rv1 is a human prostate carcinoma epithelial cell line derived from a xenograft that was serially propagated in mice after castration-induced regression and relapse of the parental, androgen-dependent CWR22 xenograft [58 (link), 59 (link)] (CRL-2505, American Type Culture Collection, Manassas, VA). The CWR22Rv1 prostate cancer cell line was kindly provided by Dr. James Jacobberger (Case Western Reserve University, Cleveland, OH), and are maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum and antibiotics. Cell cultures are regularly tested to ensure absence of mycoplasma. The CWR22Rv1 prostate cancer cell line expresses AR full-length with a duplicated DNA binding domain in exon 3 and AR splice variants, for example AR-v7, lacking a ligand binding domain. Thus, the CWR22Rv1 cell line displays constitutively active AR even in the absence of androgen [59 (link)]. In the ChIP-Seq study an AR antibody (PG21, 06-680, Sigma EMD Millipore, Darmstadt, Germany) was used recognizing both forms of the AR. All experimental protocols were approved by the Institutional Review Board at the University of California Merced. The study was carried out as part of IRB UCM13-0025 of the University of California Merced and as part of dbGap ID 5094 on somatic mutations in cancer and conducted in accordance with the Helsinki Declaration of 1975.

The 22Rv1 cell line (CRL-2505) was obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA) in 2013 and cultured in RPMI 1640 medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS) and 1× (100 μg/ml) penicillin/streptomycin. The CWR-R1 late subline was generated from the CWR-R1 cell line as described (19 (link),20 (link)), and cultured in RPMI 1640 medium with 10% charcoal stripped (CS) FBS and 1× penicillin/streptomycin. The LNCaP95 cell line was provided by Jun Luo (John Hopkins University) in 2012, and cultured in phenol red-free RPMI 1640 medium (Invitrogen) supplemented with 10% FBS and 1× penicillin/streptomycin. The VCaP cell line (CRL-2876) was obtained from ATCC in 2011, and cultured in DMEM medium (Invitrogen) + 10% FBS and 1× penicillin/streptomycin.

CWR22Rv1 is a human prostate carcinoma epithelial cell line derived from a xenograft that was serially propagated in mice after castration-induced regression and relapse of the parental, androgen-dependent CWR22 xenograft26 (link)27 (link) (CRL-2505, American Type Culture Collection, Manassas, VA). The CWR22Rv1 prostate cancer cell line was kindly provided by Dr. James Jacobberger (Case Western Reserve University, Cleveland, OH), and are maintained in RPMI 1640 medium supplemented with 10% FBS and antibiotics. Cells are regularly tested to ensure that they are mycoplasma-free. All experimental protocols were approved by the Institutional Review Board at the University of California Merced. The study was carried out as part of IRB UCM13-0025 of the University of California Merced and as part of dbGap ID 5094 on somatic mutations in cancer and conducted in accordance with the Helsinki Declaration of 1975.