Figure 1.

eNOS mRNA-targeting miR-31-5p is up-regulated in TNFα-stimulated endothelial cells. A, HUVECs were treated with TNFα (10 ng/ml) for 24 h, followed by analysis of miRNA expression profiles using Affymetrix miRNA microarrays (n = 3). Ctrl, control. B, computational target prediction using TargetScan showed complementarity between miR-31-5p and the 3′-UTR of human eNOS and generation of its 3′-UTR mutant (mt). Wildtype (WT) and mutant eNOS mRNA 3′-UTRs were cloned into the psiCHECK-2 vector. C, time-dependent expression of miR-31-5p in HUVECs stimulated with TNFα was determined at the indicated time periods by qRT-PCR (n = 5). ***, p < 0.001.