context of biological processes, pathways, and networks. Eight different temporal expression patterns identified in the analysis were 15060526 derived by combining the sign of the log2 FC variable from paired comparisons between infarct core versus remote myocardium at three temporal analyzed stages. Microarray data have been deposited in the Gene Expression Omnibus database with series accession number GSE34569. Quantitative real-time PCR To confirm microarray data, ten of the de-regulated genes in the infarct core tissue with porcine cDNA sequences commercially available, were selected for further validation by quantitative realtime PCR. Random 
hexamers and the ATL 962 cost ScriptTM OneStep RT-PCR Kit were used to synthesize first strand cDNA for each sample from 2 mg of RNA. 2 ml of cDNA were amplified in a final volume of 50 ml containing 25 ml TaqManH 26 Universal PCR Master Mix and 2 ml of each porcine FAMlabelled TaqManH Gene Expression Assay. The used primers are listed in Tissue collection and RNA extraction After sternotomy, hearts were immediately excised and washed in ice-cold buffered saline solution to remove any residual blood. Biopsies from infarct core, remote myocardium, and control myocardium were selected and immediately collected in Allprotect Tissue Reagent at room temperature to ensure RNA stabilization of harvested tissue samples. RNeasy Fibrous Tissue Mini Kit was used to isolate total RNA from tissue. RNA purity and integrity, assessed both by spectrophotometry and nanoelectrophoresis, were optimal for microarray experiments. Microarray gene expression analysis Microarray expression profiles were obtained using the GeneChipH Porcine Genome Array. Briefly, 200 ng of total RNA from each sample was processed, labeled, fragmented, and hybridized to GeneChipH according to the Affymetrix GeneChipH 39 IVT Express Kit User Manual. Washed and stained arrays were scanned using an Affymetrix GeneChipH Scanner 3000 7 G, generating. CEL files for each array. Raw expression values obtained from. CEL files for each array were Results MI alters gene expression in the whole heart In order to analyze changes in gene expression caused by infarct injury in the whole heart, microarray expression profiles of noninfarcted remote myocardium or infarct core samples at each temporal post-MI stage were compared with Gene Expression in Porcine Myocardial Infarction microarray expression profile of control myocardial samples from healthy animals . A total of six lists of differentially expressed probe sets were obtained for all comparisons. A compressed view of the cluster analysis of all expressed 19276073 probe sets showing relative differential expression in infarct core and remote myocardium at 1, 4, and 6 weeks is shown by the heat map. In this hierarchical clustering algorithm, all the 9 infarct core and all the 9 remote myocardial samples formed two distinct clusters, separately of the control samples cluster, corresponding to physiological condition. In addition, except one remote myocardial specimen from 6 weeks group, the samples also clustered by temporal post-MI stage. A total of 8,619 differentially expressed probe sets, which correspond to 6,108 unique annotated HGNC genes, were detected in the infarcted porcine heart. Fig. 2B shows the number of transcripts differentially expressed by the infarct core and remote myocardium at each temporal analyzed stage. Venn’s diagrams show overlapped regulated probe sets between both tissue regions diminishing over tim
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