Fig. 6.

miR-9 and TNF-α-mediated signaling and endothelial-mesenchymal transition (EndMT) in LECs. A: representative Western blots showing expression of VE-cadherin, N-cadherin, endothelial NO synthase (eNOS), and phosphorylated eNOS in LECs transfected with control miR, miR-9 mimics, and inhibitors. Values from 3 independent experiments were quantified and are shown below each blot. B: representative Western blots of protein samples from LECs exposed to TNF-α (20 ng/ml) for 2, 24, and 96 h showing corresponding levels of β-catenin, ZEB1, VE-cadherin, and N-cadherin. Results from experiments carried out in triplicates were quantified and are shown below each blot. Values are means ± SE. *P < 0.05. C: bromodeoxyuridine (BrdU) assay for cell proliferation. LECs were treated with miR-9 mimics or inhibitors (100 nM) in the absence or presence of TNF-α (20 ng/ml) for 2 h, and relative absorbance was measured and plotted. Experiments were done in triplicates. D: XTT assay for cell viability. LECs were treated as described in C, and XTT was added for 4 h. Relative absorbance was measured and plotted. All experiments were done in triplicates. Values are means ± SE. *P < 0.05 vs. control miR. #P < 0.05 vs. miR-9 mimic.