Recombinant and endogenous ZNF804a is localized to the nucleus of neural progenitor cells.

<p><b>A</b>) pCAG-ZNF804A was transfected into rat neural progenitor cells for twenty-four hours and processed for immunocytochemistry with antibodies against an myc-tag fused to ZNF804a. Expression of ZNF804a co-localizes with the nuclear counter stain DAPI (Scale bar = 10 µm). <b>B</b>) Immunohistochemistry using anti-ZNF804a antibodies showing endogenous ZNF804a protein (left panel) co-localizes (right panel) with the nuclear counter stain TOPRO (middle panel) in E11 neural progenitor cells within the ventricle zone. <b>C</b>) Endogenous ZNF804a protein is devoid of the cytoplasmic fraction and observed in the nuclear fraction. As a control to demonstrate proper cellular fractionation, Nestin, a cytoplasmic marker of neural progenitor cells, is observed in the cytoplasmic fraction. Likewise, Histone H3, a chromatin marker, is observed in the nuclear fraction.</p

ZNF804a regulates the transcription of several Schizophrenia- associated genes.

<p><b>A</b>) Quantitative RT-PCR was performed on 37 SZ associated gene transcripts following ZNF804A transfection. PRSS16 and COMT (red) showed robust upregulation of transcription twenty-four hours after transfection (Bonferroni corrected p<0.05; n = 5). Conversely, PDE4B and DRD2 (green) transcript levels were downregulated following ZNF804a transfection (Bonferroni corrected p<0.05; n = 5).</p

Endogenous ZNF804a binds to the DNA regions directly upstream of the transcription start site of PRSS16 and COMT.

<p><b>A</b>) Tiling qRT-PCR of the promoter sequences of PRSS16 and COMT following ChIP using anti-ZNF804a reveals enrichment for ZNF804A. Enrichment of ZNF804A on the PRSS16 promoter appears 1.5 Kb 5′ upstream of the transcription start sites (TSS) while enrichment on the COMT promoter appears 1 Kb 5′ upstream of the TSS.</p

Recombinant ZNF804a binds to the DNA regions directly upstream of the transcription start site of PRSS16 and COMT.

<p><b>A</b>) Chromatin was immunoprecipitated with antibody directed against the myc-tag or IgG as control and then probed with anti-ZNF804a antibody. Recombinant ZNF804a was correctly identified by anti-ZNF804a antibody. <b>B–C</b>) Tiling qRT-PCR of the promoter sequences of PRSS16 and COMT following ChIP against the myc-tag reveals enrichment for ZNF804A. Enrichment of ZNF804A on the PRSS16 promoter appears 1.5 Kb 5′ upstream of the transcription start sites (TSS) while enrichment on the COMT promoter appears 1 Kb 5′ upstream of the TSS <b>D</b>) Tiling qRT-PCR of the promoter sequences of DRD2 and PDE4B following ChiP did not result in enrichment of ZNF804a (all figures *p<0.05, <i>Students</i> t-test, Error bars indicate ± SEM).</p