Fig. 4: Elevated ERG and AR correlates with higher DLX1 levels representing advanced-stage aggressive disease.

a Heatmap showing TCGA-PRAD RNA-Seq data for ERG and DLX1 expression in primary PCa specimens (n = 498). Shades of red and blue represent the log₂ (norm_count+1) value. b Representative core of PCa tissue microarray (TMA) showing RNA in situ hybridization (RNA-ISH) scoring pattern for DLX1 in 144 PCa patient specimens, score 0 represents DLX1 negative, score 1 signifies low DLX1, score 2, and score 3 represents medium and high DLX1 expression, respectively. Scale bar, 50 µm. c Bar plot showing percentage of patients negative (DLX1–) and positive (DLX1+) for DLX1 expression based on the scoring pattern (P < 0.0001). d Same as b except Immunohistochemistry (IHC) for ERG (top panel) and RNA-ISH for DLX1 (bottom) in 144 PCa patient tissue specimens. e Bar plot showing percentage of patients with varying DLX1 expression in ERG-positive (ERG+) and -negative (ERG–) PCa cases (P < 0.0001). f Same as e except an association between ERG and DLX1 expression status and Gleason scores of PCa patients (P = 0.0003). g Same as b except representative tumor cores showing IHC for AR, ERG, and RNA-ISH for DLX1 representing AR+/ERG+/DLX1+ status in 144 PCa patient tissue specimens. h Same as g except for representative AR + /ERG−/DLX1+ patient in TMA containing 144 PCa specimens. i Bar plot depicting percentage of patients with positive and negative DLX1 expression in AR + /− and ERG + /− respective background. Patients showing low, medium, and high DLX1 expression categorized as DLX1-positive (P < 0.0001). j Contingency table for the AR and DLX1 status in TMA patient specimens. P-value denotes Fisher’s exact test (P < 0.0001). k Bar plot showing association between DLX1 expression and Gleason scores of tumor specimens (P < 0.0001). l Same as k, except association of DLX1 expression with tumor stage (P = 0.0041). m Correlation plot of AR and DLX1 using Stand Up To Cancer (SU2C) dataset by categorizing patients as TMPRSS2-ERG positive (top panel) and negative samples (bottom panel). P-value was calculated using two-tailed test with 95% confidence interval (P < 0.0001). n Heatmap showing DLX1 levels in tumor specimens representing distant metastatic sites of metastatic CRPC patients. o Same as n except for RNA-ISH for DLX1 expression in TMA containing 121 mCRPC biospecimens collected from various metastatic sites. Scale bar, 25 µm. p Bar plot showing DLX1 expression in percent metastatic sites from CRPC patients same as n (P < 0.0001). For panels d, g, and h scale bars are represented as 300 µm for the entire core and 50 µm for the inset image. For panels, c, e, f, i, k, l, and p P-value were calculated using Chi-Square test. Source data are provided as a Source Data file.