Fig. 5: DLX1 is a transcriptional target of ERG in TMPRSS2-ERG positive prostate cancer.

a Chromatin immunoprecipitation-sequencing (ChIP-Seq) data depicting ERG enrichment at the DLX1 promoter in the indicated cell lines from GEO databases (GSE98809, GSE110655, and GSE37752). b Schema showing chromosomal location of the ERG binding motifs (EBM) onto DLX1 promoter selected for ChIP-qPCR (top panel). ChIP-qPCR data showing recruitment of ERG, histone H3 lysine 9 acetylation (H3K9Ac), and RNA-polymerase II (RNA-Pol II) at the DLX1 and PLAU promoters. c Q-PCR (left panel) and immunoblot (right panel) data showing the expression of ERG and DLX1 in RWPE1 cells overexpressing ERG (P < 0.0001). d Schema showing site-directed mutagenesis of DLX1 promoter cloned upstream of luciferase gene, nucleotides in red were mutated (top). Luciferase reporter assay indicating wild-type (WT) and mutant (Mut) DLX1 promoter-driven reporter activity (bottom panel) in ERG overexpressing and control RWPE1 cells (P < 0.0001). e Q-PCR data showing relative expression of target genes in VCaP cells stimulated with 10 nM R1881 at the indicated time points. f Same as e, except immunoblot data. g Schematic diagram depicting ERG-mediated transcriptional regulation of DLX1 and plausible role of AR in DLX1 regulation. h ChIP-Seq data (GSE28951) showing recruitment of AR and ERG on the DLX1 enhancer and promoter regions respectively, in R1881-stimulated VCaP cells. i ChIP-Seq data (GSE70079) showing enrichment of AR at the DLX1 putative enhancer in the normal prostate (n = 6) and PCa (n = 13) tissue specimens. KLK3 represents positive control. Data shown from three biologically independent samples (n = 3). Data represent mean ± SEM. For panels, b Unpaired two-tailed Welch’s t-test; c and d Two-way ANOVA Sidak’s multiple comparison test; e Two-way ANOVA, Dunnett’s multiple comparison test was applied. Source data are provided as a Source Data file.