title: GSM9239594: Fibroblasts, extracted and cultured,BuOE; Mus musculus; RNA-Seq
library_construction_protocol: Mice were sacrificed and prostates and bladders were removed immediately post-mortem. Organs were briefly minced using a scalpel in Hank's buffered saline solution (HBSS) without calcium and magnesium, and fragments were centrifuged for 3 minutes at 400xg. Prostate or bladder fragments were resuspended in type I collagenase (Sigma-Aldrich, Saint Louis, MO, SCR103) in HBSS with calcium and magnesium and incubated at 37°C. Every five minutes, fragments were mixed by serially pipetting through a 25 ml, 10 ml, 5 ml, and 2 ml pipette until fragments could pass easily through the 2 ml pipette tip. Fragments were washed twice in HBSS without calcium and magnesium and then cultured using DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and 1% non-essential amino acids. RNA was isolated using the Qiagen RNeasy mini kit as per protocol (cat. no. 74104). RNA sequencing was performed by City of Hope (Duarte, CA). Bulk RNA sequencing was performed on the NovaSeq 6000 (Illumina, San Diego, CA) with S4 reagent kit v1.5