Quantitative profiling of differentially expressed miRNAs in siCTL- and siOASL-transfected HUVECs

Purpose: The goal of this study is to compare endothelial small RNA transcriptome to identify the target of OASL under basal or stimulated conditions by utilizing miRNA-seq. Methods: Endothelial miRNA profilies of siCTL or siOASL transfected HUVECs were generated by illumina sequencing method, in duplicate. After sequencing, the raw sequence reads are filtered based on quality. The adapter sequences are also trimmed off the raw sequence reads. rRNA removed reads are sequentially aligned to reference genome (GRCh38) and miRNA prediction is performed by miRDeep2. Results: We identified known miRNA in species (miRDeep2) in the HUVECs transfected with siCTL or siOASL. The expression profile of mature miRNA is used to analyze differentially expressed miRNA(DE miRNA). Conclusions: Our study represents the first analysis of endothelial miRNA profiles affected by OASL knockdown with biologic replicates. HUVECs were cultured in EGM-2 BulletKit medium containing 10% FBS, 2mM L-glutamine, and penicillin/streptomycin and used at passage 7-10 for experiments. Two replicates of siRNA-transfected HUVECs per group were treated with or without TNFα and IFNγ added in serum-free EGM-2 for 3hours. Endothelial micro-RNA profiles of siCTL- and siOASL-transfected HUVECs were generated by miRNA-seq.