Fig. 1: High DLX1 expression associates with poor prostate cancer prognosis and promotes disease progression.

a Dot plot showing DLX1 expression in PCa patients (n = 498) and matched normal (n = 52) in TCGA-PRAD RNA-Seq dataset, data represents log2 (norm_count+1), center depicts mean ± (standard deviation) SD (P < 0.0001). b Dot plot of DLX1 expression using microarray profiling data (GSE35988) comprising benign (n = 28), localized (n = 59), and mCRPC (n = 35) patient specimens, data represents log2 (norm_mRNA), center depicts mean ± SD (P < 0.0001). c Kaplan–Meier plot showing survival probability in TCGA-PRAD (n = 498) dataset categorized in high DLX1 (DLX1^Hi) and low DLX1 (DLX1^Lo) expression. d RNA-Seq data showing DLX1 transcript read counts in publicly available datasets (GSE128399 and GSE118206). e Cell proliferation assay using isogenic RWPE1 cells overexpressing DLX1 at indicated time-points (P < 0.0001). f Foci formation assay using same cells as e (P < 0.0001). g Boyden Chamber Matrigel migration assay using same cells as e (P < 0.0001). Representative images for panels f (scale bar 500 µm) and g (scale bar 100 µm) are shown as inset. h Cell proliferation assay using 22RV1-DLX1-KO (C-1, C-2, and C-3 are independent clones) and control cells at indicated time-points (P < 0.0001). i Boyden Chamber Matrigel migration assay using same cells as h (P < 0.0001). j Anchorage-independent soft agar assay using same cells as h (P < 0.0001). Representative images for panels i and j are shown as inset (scale bar 100 µm). k DAVID analysis showing upregulated (right) and downregulated (left) biological processes in 22RV1-DLX1-KO against control cells. Bars represent the −log₁₀ (P-value) and the frequency polygon (black line) denotes number of genes. l Same as k, except gene set enrichment analysis (GSEA) plots representing deregulated pathways. m Heatmap displaying downregulated genes involved in cancer stemness and EMT in 22RV1-DLX1-KO cells compared to control. Data shown from three biologically independent samples (n = 3). Data represent mean ± SEM unless specified. For panels, a Unpaired Student’s two-tailed t-test was applied; b One-way ANOVA with Dunnett’s multiple comparison test was applied; e Two-way ANOVA Sidak’s multiple comparison test; f, g Unpaired two-tailed Welch’s t-test; h Two-way ANOVA, Dunnett’s multiple comparisons test; i, j One-way ANOVA, Dunnett’s multiple comparisons test was applied. Source data are provided as a Source Data file.