RealTime quantitative PCR of cardiac at risk area samples obtained from a mouse model of ischemia and reperfusion injury

Mice were exposed to cardiac ischemia/reperfusion injury and sacrificed after 1 or 4 days. Hearts were stained with Evans blu/TTC to isolate the at risk area. Hearts were divided into 1mm slices, the at risk area of any slice were pooled together before RNA extraction. Mice were divided into 3 different groups: Sham animals have been subjected to the surgical procedure without the induction of the ischemia and reperfusion injury and represent the control animals; LAD group is represented by animals subjected to the surgical procedure and the induction of the ischemia and reperfusion imjury; MLAD group is represented by animals subjected to the surgical procedure and the induction of the ischemia and reperfusion injury, followed by myriocin treatment (SLN/Myr) at the beginning of reperfusion. qPCR gene expression profiling. Samples were obtained from 5 to 7 animals per group (Sham; LAD; MLAD). For any given target gene, the average expression value of all the sham animals has been compared to the expression value of each animal from LAD and MLAD group to calculate their relative expression levels. Statistical analyses were performed comparing: LAD vs Sham; MLAD vs Sham; LAD vs MLAD at both time points of reperfusion.