Flow cytometry for cell cycle analysis

Transfected AGS and BGC823 cells were washed with PBS and fixed in 75% ethanol overnight at 4°C. Cells were subsequently washed three times with PBS and stained with 500 µl eBioscience™ propidium iodide staining solution (cat. no. 00-6990-50; Thermo Fisher Scientific, Inc.) at room temperature for 15 min. Cell cycle distribution was examined using a FACScan flow cytometer (BD Biosciences) and BD Accuri C6 system software (version 1.0; BD Biosciences).