Cell cycle analysis by flow cytometry

At 72 h after transfection with miR-711 mimics, miR-711-inhibitors and empty vector, these transfected and untransfected control cells were trypsinized, resuspended in PBS and then fixed in cold ethanol at 4°C overnight. Subsequently, these treated cells were stained with 40 μg/ml propidium iodide and 100 μg/ml RNase A in PBS for 30 minutes at 37°C. Finally, the cell cycle was analysed with a BD Biosciences FACSCalibur flow cytometer.