Fig. 5. Intratumoral heterogeneity within full-tumor section 15-096M1.

a Hematoxylin and eosin (H&E) staining of 15-096M1 lymph node metastases. Distinct areas of morphology are demonstrated, cribriform well differentiated prostatic adenocarcinoma (lower left) and undifferentiated high-grade carcinoma (upper right). N = 1 tissue section for H&E staining. b Fluorescent labeling of 15-096M1 lymph node metastases. High PanCK staining is present in the lower left and low PanCK staining is present in the upper right. N = 1 tissue section for fluorescent labeling. c Individual tumor region of interest (ROIs) (200–500 µm) with varying levels of PanCK intensity and differential tumor morphology. N = 1 tissue section for fluorescent labeling and ROI selection. Expression plots of genes known to be associated with AR+/NE− (d), AR−/NE− (e), and AR−/NE+ (f) phenotypes of ROIs 7–12 from 15-096M1. Counts were Q3 normalized and scaled (Z-score) to enable plotting of all genes on the same axes. g–k Comparison of transcript levels of specific genes in ROIs 1–6 (N = 6) from the CK+ tumor region and ROIs 10–12 (N = 3) distant from the CK+ region. Counts were log₂ Q3 normalized. Significance was determined by two-sided Wilcoxon-rank tests (g–k: p = 0.024). Boxes represent the median and interquartile range (IQR) and the upper and lower whiskers extending to the values that are within 1.5 × IQR; data beyond the end of the whiskers are outliers and plotted as points.