Snu423

The human HCC cells were maintained in DMEM (Invitrogen, Grand Island, NY) with 10% fetal bovine serum (FBS), 1% Glutamine, and 1% penicillin/streptomycin. NK-92MI cells (ATCC, Manassas, VA) were maintained in α-MEM (Invitrogen) with 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, horse serum to a final concentration of 12.5% and FBS to a final concentration of 12.5% based on ATCC guidelines. All cell lines were cultured ina5%(v/v) CO2 humidified incubator at 37 °C The SK-Hep1 (ATCC, Manassas, VA) and SNU423 (ATCC, Manassas, VA) AR stable transfectants were established based on a previous procedure [14 (link)]. Cisplatin (479306), MG132 (M8699) and Cycloheximide (CHX, 227048) were purchased from Sigma.
To generate AR knock-down stable clones of SK-Hep1 (ATCC, Manassas, VA) and SNU423 cells (ATCC, Manassas, VA), HEK-293T cells were transfected with lentiviral vectors, pLKO1-sh-AR/pLKO1-scr, with the psAX2 packaging plasmid, and pMD2G envelope plasmid for 48 hrs to obtain the lentivirus supernatant, which was frozen at −80 °C for later use.
For the luciferase reporter assay, cells were transfected using Lipofectamine 3000 (Invitrogen) reverse transfection protocol, according to the manufacturer's instructions. See Supplemental data for detailed sequence information.

Human HCC and PCa cell lines HepG2, PLC/PRF/5, SNU-423, VCaP, 22Rv1, DU145 and the immortalized normal liver cell line THLE2 were obtained from American Type Cell Culture Collection (ATCC, Manassas, VA). For reproducibility experiments, HCC cells panel (HepG2/C3A, PLC/PRF/5, SNU-423, SNU-475) was obtained from ATCC (ATCC® TCP-1011). The read pairs covering exon 4–8 of the AR locus from SNU-475 cell whole genome sequencing are summarized in Supplementary Figure 1A (details in Supplementary Methods). HCCLM3 cells were kindly provided by Thomas Schmittgen, University of Florida. Cell lines have been tested and authenticated by Genetica Cell Line Testing (Burlington, NC) using short tandem repeat DNA profiling. All cells were tested for mycoplasma periodically using MycoAlert™ PLUS Mycoplasma Detection Kit (LT07–705, Lonza, Allendale, NJ) and were negative. Cells were sub-cultured in ATCC recommended medium, DMEM or RPMI-1640 (11995–065, 11875–093, Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (F0926–500ML, Sigma-Aldrich, St. Louis, MO) at 37°C in a humidified atmosphere with 5% CO₂. Phenol-red free media containing 5% charcoal-stripped fetal bovine serum (csFBS) were used for studies involving steroid hormone signaling (21063–029, 11835–030, Invitrogen, Carlsbad, CA).