Flow cytometry for cell cycle & apoptosis

For cell cycle analysis, cells were serum starved 12 hours for synchronization, re-stimulated with complete medium supplemented with 2 μM cisplatin for 24 h, fixed with 70% ethanol, and prepared for flow cytometry analysis using the Cell Cycle Detection Kit (KeyGenBiotech, Nanjing, China).

For apoptosis analysis, GC cells were seeded in 6-well plates at a density of 1.5 × 106 cells/ml. Cells were treated with a concentration of 2 μM cisplatin for 24 h, harvested using 0.2% trypsin without ethylene diamine-tetraacetic acid (EDTA), stained according to manufacturer's instructions of the Annexin V-FITC/PI Apoptosis Detection Kit (KeyGen Biotechnology, China).