GIBCOGrand Island Biological Company (tissue culture media enterprise)
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Propidium iodide (PI) was left on cells for a total of 10 min at room temperature, thereafter the cell culture media containing PI and calcein was removed and cells were washed twice with phosphate buffer solution (PBS; GIBCO, USA) and 1 ml of fresh RPMI-1640 cell culture media (with 10% FBS) was added prior to being assayed by fluorescence microscope (ZX70, OLYMPUS, Japan).