Expression profile of multipotent basal cells and basal-derived luminal cells isolated from the tip region (last 100µm) of the ductal tree of mouse prostate at P10-P13.

To investigate the molecular reprogramming of epithelial cells of the prostate during basal to luminal differentiation in vivo, we took advantage of the spatial restriction of multipotent basal cells at the distal region (tip - 100μm) of the developing prostate at postnatal day (P)10 (Tika et al., Development, 2019). The distal tips (100μm) of P10-P13 prostate glands were manually dissected from the main ducts under the stereoscope in order to enrich for multipotent basal cells and basal-derived luminal cells. Cell populations were isolated for RNA sequencing based on EpCAM and Cd49f expression via FACS. Prostate tissue of mice at P10-P13 was microdissected under a stereoscope to separate the different lobes. The ventral lobe was used to further separate (by cutting) the tips from the main ducts. Tissues were digested and living cells were isolated using fluorescence-activated cell sorting and analysis. Sorted cells were collected in lysis buffer for RNA extraction. Extracted RNA was sequenced using the bulk RNA-seq strategy.