Figure 3.

Effect of miR-193b overexpression on proliferation and apoptosis in melanoma cells. Malme-3M, SKMEL-28, and SKMEL-5 cells were transfected with miRNA precursors (either miR-193b or negative control) for 72 hours. A: Transfected cells, incubated with BrdU, were seeded at 3500 cells per well into 96-well black plate. After 24 hours, cell proliferation rate was assessed using the cell proliferation ELISA. The data are mean ± SEM from three independent experiments, each performed in octuplicate. B: Cells were fixed, stained with Annexin V FITC and PI, and analyzed by flow cytometry. Necrotic cells in D1, necrotic and late apoptotic cells in D2, viable cells in D3, and early apoptotic cells in D4. As a positive control, Malme-3M cells were treated with Mcl-1 DsiRNA and ABT-737 as described previously.20 Graphs show representative results from one of three independent experiments. C: Malme-3M cells were fixed and treated with RNase. After PI staining, cells were analyzed by flow cytometry. Data are the mean of four independent experiments, presented as mean ± SEM. ***P < 0.001 was calculated using G1 population by independent samples t test.