Apoptosis and Invasion Assay in FaDu Cells

FaDu cells post indicated treatment were harvested and suspended in the binding buffer of ApoDETECT Annexin V-FITC Kit (Thermo Fisher Scientific Inc.). After staining with fluorescently labeled Annexin V and propidium iodide (PI) (Thermo Fisher Scientific Inc.), the cells were performed with flow cytometry analyses by FACS flow cytometer (Attune, Life Technologies, Darmstadt, Germany) according to previous study [15 (link)]. For transwell assay, the FaDu cells post indicated treatment were suspended in a serum-free medium and plated into the upper chamber of the well (Corning, Tewksbury, MA, USA). Medium containing 15% fetal bovine serum was added into the lower chamber. Twenty-four hours later, cells in the lower chamber were stained with crystal violet and photographed under the microscope (Olympus). The upper chamber was precoated with Matrigel (BD Biosciences, Bedford, MA, USA) to investigate cell invasion according to previous study [15 (link)].