Fig 1: Optimization of Flow Cytometry for Purification of Stromal and Epithelial Subtypes(A) Standard flow cytometry strategy for purification of prostate stroma and epithelial subtypes.(B) Barcoding of cells from traditional FACS gates shows breakdown of cell types within each gate.(C) Quantification of cells within barcoded FACS gates.(D) (Left) CD200 labels 93% of endothelia that CD31 labels. (Right) Podoplanin (PDPN) and CD200 separate endothelia (CD200+), fibroblasts (PDPN+), and smooth muscle (PDPN−).(E) PSCA was identified as a potential cell surface marker capable of isolating other epithelial cells after CD26+ luminal epithelia are removed.(F) scRNA-seq of modified FACS gates on a new organ donor prostate specimen is used to demonstrate the increased purity of isolated stromal and epithelial cell types compared to traditional gates in (A)–(C).These data are quantitated in (G).