Postmortem brains samples from heroin abusers and control subjects were collected at the Department of Forensic Medicine at Semmelweis University, Hungary, and the National Institute of Forensic Medicine at Karolinska Institutet, Stockholm, Sweden under the ethics guidelines approved by each institution. Heroin subjects died from heroin intoxication, were predominantly heroin users not receiving methadone or buprenorphine treatment, and were negative for human immunodeficiency virus infection. No information was known about their physical dependence. The control group had negative blood levels of opiates and other drugs of abuse. Demographic information can be found in Supplemental Table 1. Striatal tissue was dissected as previously described.(18) Due to the low abundance of nucleus accumbens (NAc) tissue, putamen tissue was used for the majority of protein experiments. Putamen punches (~200 mg) were pulverized on dry ice and stored at −80°C. For laser captured samples, striatal regions were dissected from 20μm sections from human subjects using an Arcturus XT Laser Capture Microdissection Instrument (Molecular Devices, Sunnyvale, CA).
