CD31 positive-extracellular vesicles from patients with type 2 diabetes: a miRNA signature

Comparative profiling of miRNA content within CD31+EVs comparing Ctrl and T2DM patients (5 vs 5 samples with each sample prepared from the pooled plasma of 4 subjects). Plasma samples from 4 subjects were pooled to reach 1 ml. CD31+EVs were isolated from 5 Ctrl preparations and 5 T2DM preparations. RNA was extracted with a commercial kit known to enrich small RNA species (Norgen Biotek Corporation). The same amount of RNA was converted to cDNA by priming with a mixture of looped primers using the manufacturer’s instructions (MegaPlex kit, Applied Biosystem). Nine µl of cDNA were used for mature miR profiling by a real-time PCR instrument equipped with a 348-well reaction plate and human miR Array pool A containing 367 different human miR assays in addition to selected small nucleolar RNAs and negative controls (non-human miRNAs). Only miRNAs expressed in more than one sample were included in the analysis. 2-ct of the average values of each miRNA were used to build the heatmap comparing Ctrl and T2DM with the ClustVis web tool.