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. 2018 Oct 2;293(49):18989–19000. doi: 10.1074/jbc.RA118.005197

Figure 3.

Figure 3.

miR-31-5p decreases eNOS expression and NO/cGMP production. A and B, HUVECs were transfected with 80 nm control miRNA (C), miR-31-5p mimic (M), or miR-31-5p inhibitor (I), followed by stimulation with TNFα for 24 h. eNOS mRNA and protein levels were determined by qRT-PCR and Western blotting. C, cells transfected with miR-31-5p mimic or miR-31-5p inhibitor (mi-miR-31) were stimulated with TNFα for 10 h, followed by incubation with DRB for the indicated time periods. eNOS mRNA levels were analyzed (n = 5). D, cells were transfected with or without psiCHECK-2–eNOS 3′-UTR–reporter constructs (WT or mutant) alone or in combination with control miRNA (C), miR-31-5p mimic (M), or miR-31-5p inhibitor (I), followed by stimulation with TNFα for 24 h. eNOS 3′-UTR–reporter activity was determined by luciferase activity assay. E–G, cells were transfected with miR-31-5p analogues, followed by treatment with TNFα for 24 h. Intracellular NO, total nitrite, and cGMP levels were determined by confocal microscopy, Griess reaction, and ELISA, respectively. ***, p < 0.001.