Schizophrenia-Related Gene Expression Analysis

Reverse transcription was performed using RT² First Strand Kit (#330401; SABiosciences, Frederick, MD, USA), as previously described (Agarwal et al., 2016 (link)). In brief, similar amounts of RNA (750–880 ng) were added to each reverse transcriptase reaction in order to produce similar amounts of cDNA. The cDNA was used for PCR array immediately or after storage for 2–3 h at −20°C. All experiments were conducted as pairs, with samples containing cDNA from between one and four muscles for each sample in a pair. Once the large fraction of schizophrenia-related genes had become apparent, we specifically targeted additional schizophrenia-related genes in two custom PCR arrays. Expression of a total of 417 different genes was examined; 37 of these were from two custom PCR arrays (SABiosciences), and another 380 different genes were examined on five types of Human SABiosciences arrays: Common Cytokines (PAHS-021ZC), Neurotrophins and Receptors (PAHS-031ZC), Tyrosine Kinases (PAHS-161ZC), Neurogenesis (PAHS-404ZC), and Myogenesis/Myopathy (PAHS-099ZC), according to the manufacturer’s protocol with SYBR Green/ROX quantitative PCR (qPCR) Master Mix (SABiosciences 330521). All arrays were processed on Applied Biosystems 7900HT real-time PCR Systems. Data were collected using SDS 2.4 software, applying the same baseline and threshold values for all samples.