Fig. 7: BET inhibitor alone or in combination with Enzalutamide downregulates DLX1 expression and mitigates its oncogenic properties.
a ChIP-Seq data (GSE55064) showing ERG enrichment on DLX1 promoter in VCaP cells treated with JQ1 or vehicle control for 24 h. H3K27Ac represents active promoter in untreated cells. PLAU used as a positive control. b ChIP-qPCR data showing relative ERG enrichment on DLX1 promoter in VCaP cells treated with JQ1 (0.5 µM) for 48 h (P < 0.0001). c Q-PCR data showing relative expression of target genes in VCaP cells treated with Enza (10 µM), JQ1 (0.5 µM) alone or in combination for 48 h. KLK3 used as a positive control for JQ1 treatment. d Same as c except immunoblot. β-actin was used as loading control. e Same as c except 22RV1 cells. f Same as e except immunoblot (P < 0.0001). g Cell proliferation assay in VCaP cells treated with drug conditions as mentioned in c. h Same as g except 22RV1 cells (P < 0.0001). i Same as g except 42D ENZR cells (P < 0.0001). j Boyden Chamber Matrigel migration assay in VCaP cells using same treatment conditions as c. Inset shows representative image of the migrated cells (scale bar 30 µm). k Same as j except 22RV1 cells (scale bar 30 µm, P < 0.0001). l Same as j except Enza-resistant, 42D ENZR cells (scale bar 30 µm, P < 0.0001). m Fluorescence intensity of the catalyzed ALDH substrate in VCaP cells under same treatment conditions as c. Marked windows show ALDH1+ percent cell population. n Same as m except 22RV1 cells. o Same as m except 42D ENZR cells. p Bar plot showing number of spheres formed in prostatosphere assay using VCaP under same treatment conditions as c. Inset shows representative image of the spheres formed (scale bar 100 µm). q Q-PCR data for the DLX1 expression using RNA isolated from VCaP prostatospheres. Data shown from three biologically independent samples (n = 3). Data represent mean ± SEM. For panels b, c, e two-way ANOVA, Tukey’s multiple comparison test; g, h, i One-way ANOVA, Dunnett’s multiple comparison test; j–l, p, q One-way ANOVA, Tukey’s multiple comparison was applied. Source data are provided as a Source Data file.
