miR-143 binding to the 3′ UTR of PTGS2 mRNA.

<p>A, Putative miR-143-binding site in 333 bp of PTGS2 3′ UTR construct cloned into pMIR-REPORT™ for luciferase reporter assay. B, Luciferase reporter assay of PTGS2 mRNA 3′ UTR in AECs and AMCs. Luciferase activities from AECs and AMCs transfected with 200 ng (AEC) or 500 ng (AMC) of luciferase reporter plasmid containing PTGS2 3′ UTR (pMIR-REPORT_PTGS2), 10 ng of Renilla luciferase reporter pSV40-RL, and miRIDIAN miR-143 mimic (100 nM) or miR-143 hairpin inhibitor (50 nM) or equal amounts of negative controls were measured using Dual-Lucifrerase Reporter Assay System (Promega). pMIR-REPORT was used as a control. In AECs, there is a 58.5% of decrease in luciferase activity following transfection with miR-143 mimic but transfection of miR-143 hairpin inhibitor does not alter luciferase activity. In AMCs, miR-143 mimic transfection decreased luciferase activity by 25.9% compared to the control, while inhibition of miR-143 increased luciferase activity by 46.2% compared to control. Renilla luciferase activity was used for normalization of firefly luciferase activity (n = 5). The graphs show means and SE. *, <i>p</i><0.05.