Fig. 1: Single-cell RNA sequencing (scRNA-seq) of prostatectomy tissues collected from patients diagnosed with primary prostate cancer.

A For each prostatectomy (n = 10), tissue punches were collected from each zone of the prostate as well as the tumor site (4 punches per subject). For each tissue punch, part of the tissue was processed for scRNA-seq from freshly dissociated cells, and the remaining half was frozen and sectioned for histology, IHC, and in situ hybridization. Schematic created in Biorender.com. Panel A created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. B) Representative examples of hematoxylin and eosin (H&E) staining (first two column panels) and immunohitostical PIN4 staining (third column panel) of fresh frozen peripheral zone tissue punches that were benign enriched (n = 10) or cancer enriched (n = 10). Brown marks tumor protein P63 (TP63) and high molecular weight cytokeratin expressed in basal epithelial cells. Red marks alpha-methylacyl-CoA racemase (AMACR), which is expressed in carcinoma and prostatic intraepithelial neoplasia (PIN). For 1x magnification, scale bar indicates 2.5 mm. For 10x magnification, scale bar indicates 200 μM. C Dimensionality reduction (uniform manifold approximation and projection, UMAP) and clustering analysis of scRNA-seq libraries from peripheral zone (PZ) tissue showed cells clustering by recognized cell types (n = 110,715 cells, 18 samples). Subject and cancer-enriched zones of luminal cells suggest inter-individual and intra-tumor heterogeneity in cancer cells. D Heatmap of cell type marker genes show cluster-specific expression.