Total neuronal RNA from three independently generated batches of iN cells for each cell line was prepared using TRIzol ® Reagent (Thermo Fisher Scientific), human-specific Taqman probes were purchased for OPRM1, MAP2, Tuj1, VGAT, GAD1, TH and PCR reaction conditions followed the manufacturer’s recommendations. Undifferentiated iPS cells, ES cells, and mouse astrocytes were used as negative controls. A sample of total RNA of a healthy human brain as well as human Thalamus from Biochain ® was used as a positive control. Relative RQ values were obtained by normalizing expression levels to the C12 iN condition. Student’s t-test was used to compare grouped N40 and D40 means. Evaluation of qPCR was performed blind to the genotype. See Supplemental Table 1 for a comprehensive list of primers and probes used in this study.
