Characterisation of VDR signaling in prostate cancer health disparities (RNA-Seq; BAZ1A conditional)

To investigate the effect of 1,25(OH)2D3 activation on RNA expression in prostate cell lines derived from European Americans and African Americans Overall design: GFP-BAZ1A was purchase from Addgene (plasmid # 65371)and cells were stable transducted by selection and maintenance in media supplemented with puromycin (2µg/mL).RNA was extracted from cells in the presence of 1a,25(OH)2D3 (100nM, 8hr) or EtOH in biological triplicate samples and analyzed by RNA-Seq. Sequencing libraries prepared with the TruSeq Stranded Total RNA kit (Illumina Inc), from 1ug total RNA. Alignment of raw sequence reads to the human transcriptome (hg38) was performed via Rsubread and transcript abundance estimates were normalized and differentially expressed genes (DEGs) identified using a standard edgeR pipeline. Comparative RNA gene expression profiling analysis of RNA-seq data for prostate cell lines treated with 1a,25(OH)2D3 (100 nM, 8h), with stable expression of BAZ1A or vector control