Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit

The cells (4 × 10 5 per well) were seeded in a six-well plate overnight and treated with varying concentrations of VPA for 4 days. To measure the extent of cell apoptosis, an Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit was used according to the manufacturer's instructions (Sungene Biotech, Tianjin, China). In brief, the cells were harvested and suspended in 1 × binding buffer. Then, 100 µL of the cell suspension was incubated with 5 µL of Annexin V-FITC for 10 min, followed by incubation with 5 µL of PI solution for another 5 min. The labeled cells were then assessed by flow cytometry and analyzed using Cellquest 6.0 (BD Biosciences, NJ, USA) [15] .