thermal cycle profile was programmed for denaturation for 15 s at 95°C and annealing for 60 s at 60°C. A total of 40 PCR cycles were performed. Data analysis was performed using the ΔCt method using the Data Analysis Template Excel file provided by SABioscience Corporation as published previously (Lee et al., 2008). Values for ΔCt were normalized to the average of five housekeeping genes: glucuronidase β, hypoxanthine guanine phosphoribosyl tranferase1, heat shock protein 90kDa α class b member 1, glyceraldehydes-3-phosphate dehydrogenase, and β actin. As recommended by SABioscience, a 2-Way ANOVA with Bonferroni post-tests (using average ΔCt values) was used to detect changes across the entire array as well as individual genes while accounting for multiple comparisons to reduce the probability of type II errors. The percent of expression relative to the adolescent control group (P38) is presented for each gene.
