miR-486-5p Restrains Extracellular Matrix Production and Oxidative Damage in Human Trabecular Meshwork Cells by Targeting TGF-β/SMAD2 Pathway

Figure 3

miR-486-5p targets SMAD2. (a) Twelve genes (FGF7, BTAF1, ST5, FOXO1, TOB1, SLC38A1, TWF1, ARID4B, RELT, PTEN, ARHGAP5, and SMAD2) that were overlapped among target-predicting algorithms are shown as a Venn diagram. (b) Expression of candidate targets in miR-486-5p-overexpressing TMCs was assessed by RT-qPCR. (c) Analysis of the SMAD2 3’-UTR showed a binding site for miR-486-5p. (d) In miR-486-5p-overexpressing TMCs and HEK293T cells, the luciferase activities of reporters with a wild type or mutant SMAD2 3’-UTR were detected. (e) Relative expression of miR-486-5p and SMAD2 in TMCs treated with varying concentrations of H₂O₂ for 12 h are quantified by qPCR. After TMCs were disposed with 300 μM H₂O₂ alone, 300 μM H₂O₂ plus NC mimics alone, or 300 μM H₂O₂ plus miR-486-5p mimics, (f) the SMAD2 level was determined by RT-qPCR; and (g) protein expression of SMAD2 and TGF-β was determined by western blotting. ; .

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