Annexin V-FITC/PI staining for apoptosis evaluation

Flow cytometry assay was performed following the kit guidelines. Briefly, SH-SY5Y cells were trypsinized after appropriate treatment, rinsed twice with ice-cold PBS, centrifuged at 1000 rpm for 5 min and re-suspended the cells in Annexin V-FITC/PI binding buffer (195 μL). Annexin V-FITC (5 μL) was supplemented and the cells were stored in dark at room temperature for 30 min. Cells were then pellet down by centrufing at 1000rpm for 5 min and re-suspended in Annexin V-FITC/PI binding buffer (190 μL); Propidium Iodide (PI) (10 μL) was further added, followed by incubation in the dark for 5mins. The apoptotic cells were quantified using Flow cytometer.