Fig. 1. Aberrant activation of HGF/MET and Wnt signaling pathways in human double-negative AR-null NE-null prostate cancer patients.

a Heatmap showing score of indicated gene signatures or expression profile of indicated genes across human metastatic castration-resistant prostate cancer (mCRPC) samples obtained from SU2C/PCF RNA-seq datasets (2019) at cBioPortal^12,13 (ARPC AR-active prostate cancers, n = 177; NEPC neuroendocrine prostate cancers, n = 13; DNPC double-negative AR-null NE-null prostate cancers, n = 14). Scores of AR- or NE-associated gene signature are shown in the top panel based on the previous study¹³. HGF/MET pathway-associated genes and Wnt/β-catenin downstream target genes are listed in the middle and bottom panel, respectively. Colors reflect the level of gene signature score or expression. See “Methods” section. b Gene Set Enrichment Analysis (GSEA) enrichment plots of pre-ranked gene list from differentially expressed genes (DEGs) comparing DNPC to ARPC samples. NES normalized enrichment score, FDR false discovery rate. See Supplementary Data 1. c Heatmap of pairwise Spearman correlation between the indicated gene signatures in indicated CRPC samples. Numbers indicate correlation coefficient. Colors reflect the correlation coefficient value. d Scatter plots displaying the mRNA expression z-scores of indicated gene signatures in indicated CRPC samples. The pink lines show association between the gene signatures and r indicates Spearman’s correlation coefficient. e Representative images of immunohistochemistry staining using the indicated antibodies on tumor tissues from naive primary prostate adenocarcinoma (PCa, n = 5) and androgen deprivation therapy (ADT)-treated CRPC patients (n = 6, please also see the “Material” section). AR androgen receptor, NE neuroendocrine, E-CAD E-cadherin, SYN synaptophysin, pMET phosphorylated MET, β-cat β-catenin. Representative images from three independent experiments with similar results are displayed for each micrograph. Scale bars, 25 μm.