Figure 5

miR-29 s down-regulated MMP- 2 in ARPE- 19 and EA hy926 cells. A, ARPE-19 or EA hy926 cells were cultured in six-well dishes and transfected with miR-29a, b, and c (50 nM), and NC (50 nM) individually when the cell densities reached approximately 70% confluency. After being cultured for 48 hours, the supernatants were harvested for examining the MMP-2 contents with western blot as described in Methods. The images shown are typical of those from three independent experiments with each condition from triplicate cultures. B, The ratios of Mock between supernatant MMP-2 and protein mass in culture wells were set as 1 and the values for treated conditions were normalized to the control values. C, HEK-293 cells were cultured in 96-well dishes, and each well was transfected with 50 ng pMIR-MMP-2 3’UTR/firefly luciferase, 25 ng pRL-SV40 Renilla luciferase vector, and 5 nM miR-29 mimics or 5 nM NC. Luciferase activities were determined 24 hours after transfection and the results were expressed as the ratios between the activity of firefly luciferase and that of Renillaluciferase. The results shown were mean (±SEM) from three independent experiment with each condition per experiment from quadruplicate cultures. The values of control were set as 1 and the results from treatments were normalized to control. *P < 0.05 versus NC. NC, negative control; MMP-2, matrix metallopeptidase-2; UTR, untranslated region.