FIG. 4.

Effect of miR-192/215 on E-cadherin expression in proximal tubular cells. A: Proximal tubular cells were transfected with either miR-control (miR-NC), miR-192, or miR-215 (100 nmol/l), and E-cadherin expression was assessed by real-time quantitative PCR. Both miR-192 and miR-215 resulted in an increase in E-cadherin mRNA, but only the change with miR-215 was significant (*P < 0.05, compared with control). B: To test the effect of miR-192/215 in the context of TGF-β, NRK52E cells that were previously adapted to TGF-β for >10 days were transfected with miR-control, miR-192, or miR-215, and expression of E-cadherin was assessed by real-time quantitative PCR 3 days later. Both miR-192 and miR-215 significantly increased E-cadherin mRNA (*P < 0.01, compared with control). C: Similar transfection experiments using miR-192 did not alter mRNA levels of ECM genes or vimentin, compared with miR-control. D: Transfection with miR-215 did not alter the expression of ECM genes or vimentin compared with miR-control. E: NRK52E cells that had not been previously exposed to TGF-β were transfected with miR-control, miR-192, or miR-215, and 4 h later treated with TGF-β (10 ng/ml). Cells were harvested 3 days later, and expression of E-cadherin was assessed by real-time quantitative PCR. TGF-β treatment dramatically reduced E-cadherin mRNA levels in cells transfected with miR-control (*P < 0.001, compared with no TGF-β treatment). The reduction in E-cadherin mRNA was partially but significantly reversed by both miR-192 and miR-215 (#P < 0.05, compared with miR-control with TGF-β treatment). All error bars represent ± SEM.