NextSeq 500 sequencing; RNA-sequencing of prostate cancer xenografts, their 3D cultures and prostate cancer cell-lines.
RNA-sequencing of prostate cancer xenografts, their 3D cultures and prostate cancer cell-lines.
PC3, DU-145, 22rV1, LAPC-4 and LNCaP cell lines were cultured in RPMI 1640 (21875-034 Life Technologies)supplemented with 10% Fetal Bovine Serum (FBS-11A Capricorn Scientific) and 1% Penicillin/Streptomycin(15140-122 Life Technologies) with 5% CO2 at 37°C. LAPC4 were in addition supplemented with 1 nM DHT.The LNCaP-abl cell line was cultured in Phenol-Red Free RPMI 1640 (11835063 Life Technologies) containing10% charcoal-stripped serum (CSS Fetal Bovine Serum, Charcoal Stripped A3382101 Life Technologies) and1% Penicillin/Streptomycin with 5% CO2 at 37°C.VCaP  cell lines were cultured in DMEM(61965059 Life Technologies) supplemented with 10%FBS and 1% μg/ml Penicillin/Streptomycin with 5% CO2 at 37°C.MDA-PCa-2b cell line was cultured in ATCC-formulated F-12K medium (30-2004) supplemented with 20% FBS,25 ng/ml Cholera Toxin (C8252 Sigma), 10 ng/ml Epidermal Growth Factor (AF-100-15 Peprotech), 0.005 mMPhosphoethanolamine (P1348 Sigma), 100 pg/ml Hydrocortisone (H0135 Sigma), 45 nM Selenium Acid (211176Sigma), 0.005 mg/ml Human Recombinant Insulin (I1884 Sigma) and 1% Penicillin/Streptomycin with 5% CO2 at37°C. RNA was extracted using the Rnasy Mini Kit (Qiagen) and sample quality was assessed using Agilent Bioanalyzer. RNA libraries were prepared for sequencing using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina®
Experimental Factor: growth condition
xenograft
Experimental Factor: cell line
LNCAP-ABL