MicroRNA quantification
AMI
Total RNA from ProtheraCytes and their secreted exosomes collected from 7 AMI patients were isolated respectively using a miRNeasy Tissue/cells Advanced MiniKit and a miRNeasy Serum/Plasma Advanced Kit (QiAGEN, France) according to the manufacturer’s protocols. RNA isolated from exosomes and cells was reverse transcribed to cDNA using the miCURY LNA RT Kit (QIAGEN, France). UniSp6 RNA spike-in controls were added during cDNA synthesis to ensure the quality of the experiment. Real-time qPCR amplifications were performed for each RT reaction. Reactions were performed according to the manufacturers’ instructions using a miRCURY LNA miRNA SYBR Green PCR Kit (QIAGEN, France) with the Bio-Rad CFX96 Real time PCR Detection System (BioRad Laboratories, France). All primer sets were custom designed by the supplier. Primers used were miR-21-5p (YP00204230), miR-26a-5p (YP00206023), miR-126-3p (YP00204227), miR-130a-3p (YP002046658), miR-133a-3p (YP00204788), miR-146a-5p (YP00204688), miR-199a-3p (YP00204536), miR-378a-3p (YP00205946), miR-590-3p (YP00205448), miR-24-3p (YP00204260), miR-29b-3p (YP00204679) and miR-132-3p (YP00206035), miR-34a-3p (YP00206061), miR-143-3p miRNA (YP00205992), and miR-506-3p (YP00204539). The qPCR data were normalized to miR-let7a-5p (YP00205727) values. Relative miRNA expressions were calculated using the 2