Fig. 5

MiR-200a expression suppresses renal carcinoma cell proliferation and induced cell cycle arrest at G0/G1 phase in renal carcinoma cells. a EdU proliferation assay analysis of the effect of miR-200a overexpression or inhibition on the proliferation of renal carcinoma cells. The EdU proliferation assay was performed 48 h after the transfection of ACHN cells with miR-NC, miR-200a mimics, miR-NC inhibitor, or miR-200a inhibitor, the representative images are shown and the ratio of EdU-positive ACHN cells are also shown at the right bottom. The cells with red fluorescence are in the S phase of mitosis, and the cells with blue fluorescence represent all of the cells. b, c Flow cytometry analysis of the effect of miR-200a overexpression (b) or inhibition (c) on the cell cycle profile changes of renal carcinoma cells. Fluorescent-activated cell sorting analysis were performed 48 h post-transfection with miR-NC, miR-200a mimics, miR-NC inhibitor, or miR-200a inhibitor in ACHN cells. After transfection, the cells were collected and washed once with PBS, resuspended in 300 μL of PBS, and fixed with 700 μL of 70% ethanol at 4 °C for overnight. Fixed cells were washed twice in PBS and stained with 1 μg/ml propidium iodide (PI) solution containing 10 μg/ml RNase (Sigma, USA) in PBS at 37 °C for 1 h in the dark. Stained cells were then analyzed using a flow cytometry with the FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA), and the relative cell population were shown at the right bottom. Representative experiment was performed in triplicate. Data are shown as the mean ± s.d. ^**P < 0.01; ^***P < 0.001