To examine if ZNF804a expression may directly regulate transcription, we performed ChIP to determine whether ZNF804a directly interacts with genomic regions proximal to the promoters of COMT, PRSS16, PDE4B, and DRD2, the four regulated genes indentified in our qRT-PCR assays. Chromatin samples were isolated from progenitor cultures transfected with pCAG-ZNF804a. Proteins were cross-linked to DNA prior to isolation and then sheered, chromatin was immunoprecipitated with an anti-myc antibody, crosslinks reversed, and isolated DNA subjected to qRT-PCR with a set of tiled primers spanning 2 kb upstream of the annotated transcription start sites for PRSS16, COMT, PDE4B, and DRD2 that included a GC-rich region (Table S2; PRSS16, chr17:50145344-501473440, COMT ch11:84579705-84581705; PDE4B, chr5:133600726-133602726 and DRD2, chr8: 50582050-50584050). To confirm our epitope-tagged expression construct, we immunprecipitated protein from transfected cells with anti-myc-tag antibody and detected it in western blot with an anti-ZNF804a antibody (136 kDa; Figure 3a). For our ChIP assays 1 µg of DNA was immunoprecipitated with anti-myc-tag antibody or negative control IgG and expressed as a percentage of input. Chromatin immunoprecipitated for ZNF804a-myc was enriched in single distinct peaks within the
