For validation of array data by Fluidigm technology 22 animals (88 samples) were kept to fit the technical constraints of this technique. Total RNA (1 μg) used in microarray experiments was reverse-transcribed as previously described . Primer sequences for genes were designed using Primer3plus software (http://primer3plus.com) and are given in Additional file 1. The TFRC gene (transferrin receptor) and EPRS gene (glutamyl-prolyl-tRNA synthetase) were used as internal controls. Pre-amplified samples were analyzed with a 96 ×96 Dynamic Array™ IFC (Fluidigm) following the protocol defined by , with some modifications. All measurements were performed on the same plate. Each gene was tested twice for each sample. Four dilution points containing a pool of all samples were used to determine PCR efficiency. Data were analyzed using BioMark Gene Expression Data Analysis software (Fluidigm) to obtain Ct values. The Pfaffl method was applied to compute the relative expression of each gene . Pearson correlations were computed to compare the expression values of microarray and quantitative real-time PCR. Quantitative RT-PCR data were also analyzed for time effect by ANOVA with repeated measurements for every gene.