Quantitative real time RT‐PCR (qPCR)

qPCR was performed to validate microarray results (on the same RNA samples used in the microarray) and to measure vWF expression (N: NS‐S = 6; S‐S = 6). cDNA was obtained using the High Capacity Reverse Transcription kit (Applied Biosystems, Branchburg, NJ). cDNA templates (10 ng) were processed by qPCR using the 7900HT thermal cycler apparatus equipped with the SDS software version 2.3 (Applied Biosystems) for data collection. Taqman primer sets (Applied Biosystem, Supporting Information Table S1) were used for cDNA amplification, and Ct values were normalized to measures of TBP and PGK1 mRNA (Taqman primers code TBP: Mm00446974_m1; PGK1: Mm01225301_m1). All data were run in triplicate and analyzed using the ΔΔC(t) method. Correlation analyses between microarray and qPCR expression data were performed correlating (Pearson correlation) the expression data for each animal obtained with the two techniques.