Apoptosis assay by flow cytometry

Apoptosis could be detected by staining the cells with Annexin V-FITC and Propidium iodide (PI) solution followed by flow cytometry analysis.11 In brief, cells were seeded to a density of 5×10⁵ in a six-well plate and treated with D. maritima (10, 100, and 500 µg/mL) for 48 hours. Then, cells were washed with cold PBS and re-suspended in the 1x binding buffer containing Annexin V-FITC and PI solution. The stained cells were examined by FACS Calibur flow cytometry (BD Biosciences, San Jose, CA, USA).