Extracellular Vesicle RNA Isolation and Sequencing

RNA isolation from ASC‐EVs, library generation, cluster amplification, and sequencing were performed by Macrogen (Seoul, Korea). Samples for small RNA sequencing were prepared as described previously (Jung et al., 2020 (link)). Briefly, RNA was extracted from ASC‐EVs using the Maxwell RSC miRNA from plasma (Promega). Libraries were then generated using the SMARTer smRNA‐seq Kit for Illumina (Takara Bio, Shiga, Japan) according to the manufacturer's protocol. The Illumina HiSeq 2500 instrument (Illumina, San Diego, CA, USA) was used for sequencing the libraries. Target genes of each miRNA in ASC‐EVs were predicted using publicly available algorithms, including TargetScan 7.2 (http://www.targetscan.org) and miRDB (www.mirdb.org).