RNA-SEQ analysis of the superficial, intermediate and basal cell layers of the bladder urothelium 24h after infection with UTI89 strain of E.coli and their adult female non-UTI controls.

Dissection and FACS cell sorting techniques were used to isolate the superficial, intermediate and basal cell types from the bladder urothelium of adult female mice following UTI infections, and their controls. This analysis will determine the transcriptional profile of each cell type, identify compartment specific transcripts, compartment specific transcript isoforms and cell-type specific noncoding RNAs. Overall design: Total RNA was obtained from FACS isolated superficial, intermediate and basal cell layers of the adult female bladder urothelium 24h after infection with UTI89 strain of E.coli and also from their non-UTI controls. Tg(Upk3a-GFP/cre/ERT2)26Amc x Gt(ROSA)26Sor and Tg(KRT5-cre/ERT2)2Ipc x Gt(ROSA)26Sor) transgenic mice were used to isolate the intermediate and basal cells, respectively. The long term goal is to generate a transcriptional atlas of bladder urothelium following UTI infections.