Commercial human prefrontal cortex RNA was obtained from Ambion (Austin, TX). Fifteen control and fourteen alcoholic postmortem prefrontal cortices were obtained from the Tissue Resource Centre at the University of Sydney. The samples were matched as closely as possible by age, gender, brain pH, RNA integrity number, and post-mortem interval (Table S1 in Supplement 1). Control subjects were defined as those who drink <20g/day (most were social drinkers or teetotallers). Alcoholics were defined based on alcohol consumption of >80g/day. Diagnoses were confirmed by physician interviews, hospital medical record reviews, questionnaires to next-of-kin, and from pathology, radiology, and neuropsychology reports. The Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) diagnosis was based on a clinical assessment by specially trained staff in psychiatry or psychology (17). After RNA extraction, DNase (Ambion, Austin, TX) was treated?, and ribosomal RNA (rRNA) was depleted using the RiboMinus kit (Invitrogen, Carlsbad, CA).
