MCAO-Induced Stroke: Blood-Derived RNA Analysis

The methods employed for RNA extraction and amplification were adapted from Martha et al. [8 (link)]. Blood was collected from the jugular vein at three different time points: immediately prior to MCAO surgery, 5 minutes after MCA reperfusion, and 72 hours post-MCAO procedure. Sham rats underwent the complete 5t-MCAO procedure, with the exception that the filament was not placed to induce a stroke; blood samples were taken at the same time points for sham animals. Total RNA was extracted from the pellet/buffy coat using the Nucleospin Blood Kit (Macherey-Nagel, Düren, Germany). RNA quantity was assessed with a Nanodrop Lite (Thermo-Fisher; Waltham, MA). cDNA synthesis utilized the RT² First Strand Kit from Qiagen, and gene expression analysis of 84 genes was performed using the ABI StepOne Plus Qiagen (Germantown, MD) and the RT² Profiler Rat Chemokine and Receptor Array from Qiagen.