Cell Cycle Analysis by Flow Cytometry

After 72 h transfection, the cells were obtained via centrifugation, rinsed with Phosphate buffer saline (PBS) and added with 300 μl PBS for resuspension. Then, 700 μl 100% ethanol was added into each group for cell fixation at 4°C for 24–48 h. The cells were subsequently cleaned and stained using 100 μg/ml propidium iodide solution and 50 μg/ml RNase solution (Invitrogen, USA) at 37°C for 30 min avoiding light. Finally, flow cytometry (BD) was utilized for cell detection following the removal of cell clumps; then, CELL Quest and ModFit LT softwares were utilized to analyze the result [15 (link)].