Rat gene 1.0 st

Rat Gene 1.0 ST Affymetrix arrays were used for evaluating the whole‐transcript expression profiling. Microarray analysis was performed at the INMEGEN microarrays core facility and all procedures were according to Affymetrix's protocol. Briefly, double‐strand cDNA was obtained from total RNA using Superscript II reverse transcriptase with poly(T) oligomer. Then, cDNA served as a template for generating biotin‐labeled cRNA. The GeneChip hybridization Oven 645 (Affymetrix) provided controlled temperature and rotation for hybridization for 17 h at 45°C. After hybridization, arrays were washed and stained by using streptavidin‐phycoerythrin in an Affymetrix Fluidics Station 450 (Affymetrix). Then, microarrays were scanned by GeneChip Scanner 3000 7G (Affymetrix). Data normalization was performed by using the transcriptome analysis Console 4.0 software (Robust Microarray Average algorithm; Thermo Fisher Scientific Inc.) and differential gene expression analysis (fold change cut‐off ±2, analysis of variance [ANOVA], p ≤ .05). The R ggplot2 library was used to generate plots of principal component analysis (PCA). Intergroup hierarchical clustering was performed by using IBM SPSS Statistics (IBM) using total normalized microarray expression. ClustVis software was used to construct the heat map graphic of differentially expressed genes.