GSM6265733: LNCaP, RNASeq, 6hr, DMSO, Rep 3; Homo sapiens; RNA-Seq
LNCaP cells (Clone FGC, ATCC CRL-1740) in RPMI1640 5% FBS were seeded at a density of 300,000 cells / well in 6 well plates. 16 hours later, wells were refreshed with seeding media spiked with either EPI-001 or 1ae at the doses indicated and 0.5% v/v DMSO control. After 6 and 24 hours, media was removed and cells were harvested using 300 µL of TRIzol reagent (Thermo 15596026) for each well. RNA was then extracted using a Zymo DirectZol Micro kit (Zymo R2062) according to the manufacturer's protocol. Total RNA-seq libraries were then prepared using 1 µg of RNA from each sample and the KAPA RNA HyperPrep Kit with RiboErase (Roche KR1351) according to the manufacturer's protocol with 10 amplification cycles. Libraries were sequenced on a NovaSeq 6000 with paired-end reads of 100 base pairs, with a read depth of 50 million fragments / library. Three libraries from three corresponding biological replicates  were prepared for each treatment (time, dosage, and compound).