Figure 3. Functional effects of miR-493 in transfected T24 and J82 cells.

A. MiR-493 expression levels in bladder cancer cell lines (T24, J82) were detected by using real-time PCR at 48 hr after transfection of miR-493 precursor. Data are presented as mean value ± SD for three independent experiments and compared to the level of miR-493 in SV-HUC-1 cells normalized as 1. B. Cell viability was analyzed by the MTS assay 4 days after transient transfection. C. Wound healing assay with miR-493 transfected cells. At 48 hr after transfection, cells were transferred from 6-well to 24-well plates and further incubated for 24 hr. A wound was formed by scraping and the wound measured after 24 hr (T24 cells) and 72 hr (J82 cells). D. Migration assay with miR-493 transfected cells. At 72 hr after transfection, cells were added into the chamber. Cells were incubated for 4 hr at 37°C in a 5% CO2 tissue culture incubator, no migrated cells were removed from transwell membrane filter inserts using cotton-tipped swab and migrated cells were stained with Hema 3 STAIN SET. Representative photomicrographs are shown at 100x magnification. E. MiR-493 and miR control were transfected into T24 and J82 cells. At 72 hr after transfection, cell cycle assay was carried out by flow cytometry.