Figure 7. Defined ERG target gene signature is not increased in 1,25D(OH)₂D₃-treated VCaP cells.

(A) LNCaP or VCaP cells were changed to stripped serum and treated for 24 hours with vehicle (EtOH), 10 nM 1,25D(OH)₂D₃ (1,25D), 100 nM 1,25D(OH)₂D₃, or 10 nM R1881 and RNA was purified. ERG mRNA was measured by RT-qPCR and normalized to 18S. (B) Venn diagram showing comparison of a previously published microarray from VCaP cells treated with siRNA against ERG (siERG IVB), a previously published microarray from VCaP cells treated with DHT, and our RNA-seq data from VCaP cells treated with 10 nM 1,25D(OH)₂D₃. The siERG IVB group denotes genes significantly downregulated at least 1.5-fold relative to control siRNA (genes requiring ERG for expression) and the DHT and 1,25D(OH)₂D₃ groups denote genes significantly upregulated at least 1.5-fold relative to vehicle control. (C) The top 10 pathways significantly regulated via GSEA using the Hallmark gene set collection comparing siERG IVB (ERG induced),1,25D(OH)₂D₃, and VDRM2-treated VCaP cells plotted as normalized enrichment score. *p < 0.05, ***p < 0.001, relative to respective LNCaP treatment. n = 3, representative graph, mean ± SEM.