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LNCaP human prostate cancer cells (American Type Culture Collection; Bethesda, MD) were maintained in RPMI-1640 media (StemCell Technologies; Vancouver, BC) supplemented with 10% fetal bovine serum (FBS; StemCell Technologies) and incubated at 37C with 5% CO2. Cells at passage 38 were plated at a density of approximately 4x10^6 cells per T175 flask. Cells were serum-starved for 48 hours prior to treatment for 16 hours with 10 nM R1881 (PerkinElmer; Woodbridge, Canada). Cells were harvested and total RNA was extracted from the cells using TRIZOL Reagent (Invitrogen Life Technologies, Carlsbad, CA) following the manufacturer's instructions
Nucleotide
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