Transcripts identified as differentially expressed in microarray analyses were independently assayed by quantitative real-time RT-PCR using TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA): IL8 (Hs00174103_m1), CDC25B (Hs00244740_m1), IL1B (Hs00174097_m1), EGR1 (Hs00152928_m1), FOSB (Hs01547109_m1), PTGS2 (Hs00153133_m1), IFI6/G1P3 (Hs00242571_m1), GAPDH (Hs99999905_m1). We also verified microarray indications of equivalent expression of GR mRNA, NR3C1 (Hs00353740_m1). Assays for each sample (n = 14) were carried out in triplicate using an iCycler instrument (Biorad, Hercules, CA, USA), Quantitect Probe RT-PCR enzymes (Qiagen), and the manufacturer's recommended 1-step thermal cycling protocol. ISGF12 was assayed using published primer sequences and reaction conditions [33]. Threshold cycle numbers for each analyte were normalized to GAPDH for analysis. A multivariate analysis of variance (MANOVA) tested group differences among all analytes simultaneously, and independent sample t-tests assessed differential expression of each individual transcript.
