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Chunk #1 — Methods — Human Tissue

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Expression of ZNF804A in human brain and alterations in schizophrenia, bipolar disorder, and major depressive disorder: a novel transcript fetally regulated by the psychosis risk variant rs1344706.
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Gray matter was dissected out, snap frozen, and stored at −80°C prior to RNA extraction, which was carried out using standard methods. The DLPFC tissue from this series was also used for protein extraction and Western blotting. The second brain series, used for immunohistochemistry, came from the Stanley Medical Research Institute, who provided free-floating sections of inferior parietal lobe from men with schizophrenia and male control participants (eTable 1 in the Supplement).30 For this series, after brief formalin fixation of the cerebral hemisphere, 3- to 5-cm coronal slabs were cut, formalin fixed for 3 to 15 weeks, and stored in buffered 30% sucrose. All blocks containing inferior parietal lobe were serially cryostat sectioned at 60 μm. Participants from both series were genotyped for rs1344706 using TaqMan assay C_2834835_10 (Life Technologies). Immunohistochemistry was also carried out on slide-mounted sections cut from formalin-fixed, wax-embedded blocks of frontal, temporal, or visual cortex from 5 additional adults and of frontal cortex from 2 neonates. The NIMH/Lieber Institute for Brain Development brains were collected with audiotaped oral informed consent from legal next of kin under