title: GSM8370725: G2S9naive2; Mus musculus; RNA-Seq
library_construction_protocol: tissue procurement: Prosates from naïve or inflamed POET3 mice Minced mouse prostate tissues were digested in 1 mg/mL collagenase (Sigma-Aldrich) in RPMI-1640 (Gibco) media containing 10% FBS (Corning) with shaking at 37ºC for 2 hours, followed by trypsinization. Dissociated cells were passed through 20G needles and 40 µm cell strainers to eliminate aggregates, followed by removal of red blood cells by ACK buffer. To enrich basal prostate stem cells (bPSC), isolated cells were stained with Zombie Violet Live/Dead Fixable Viability Dye (Biolegend) in PBS, followed by incubation with 1:100 diluted fluorescence-conjugated specific antibodies (Biolegend): CD45-FITC (#103108), CD31-FITC (#102506), Sca-1-APC (#122512) and CD49f-PE (#313612). Once stained, fluorescence activated cell sorting was performed on the BD FACSAria under sterile conditions. Data was collected using the C1 Fluidigm instrument with SMARTer chemistry (Clontech) to generate cDNA from captured single cells. Libraries were prepared using a Nextera kit (Illumina). Single-end 1x50 bp reads were sequenced using the HiSeq2500 on rapid run mode.