For quantitative RT–PCR analyses of pooled cultured cells, RNA was isolated using the RNAqueous Kit (Applied Biosystems), treated with DNase (Applied Biosystems), and reverse-transcribed with Superscript III (Invitrogen). mRNA levels were quantified by real-time PCR assay using the Applied Biosystems 7900HT Fast real-time PCR system and RQ analysis software. For quantitative RT–PCR analyses of single cells, cytoplasm from individual cells was aspirated with a patch pipette, and mRNA levels were measured in the cytoplasm using the Fluidigm Biomark dynamic array system as described (Pang et al., 2011). For all quantitative RT-PCR assays, titrations of total human brain RNA were included in each experiment, and only primers that demonstrated a linear amplification with R2 values of >0.98 were included (see Supplemental Materials and Table S1 for details).
