Quantitative Real-Time PCR (qRT-PCR) was used to confirm differences in 4 genes: FKBP5, GRM3, NR3C1, and NR4A2. Primers were selected from Life Technologies™ (Carlsbad, CA) catalog of Taqman® Gene Expression Assays (http://bioinfo.appliedbiosystems.com/genome-database/gene-expression.html). One µg of total RNA from each sample was used for reverse transcription using the High Capacity cDNA Reverse Transcription Kit (Life Technologies™, Carlsbad, CA). Each gene of interest was measured in duplicate using TaqMan® Fast Advanced Master Mix (Life Technologies). Primers for POL2RA (Taqman® primer: Hs00172187_m1) were included in each well as a control. The CT of the POL2RA run in the same well was subtracted from the CT of the target gene to yield the Delta CT (relative expression). The Delta CT from 2 replicates for each sample was used in a 3-way ANOVA using phenotype, sex, and sample ID as factors.
