Flow cytometry of sunitinib-tolerance 786-O cell

Cells (2×10⁵) were cultured in 6-well plates, and then were incubated with sunitinib for 24 hr. Necrosis was analyzed using the Annexin V-FITC/PI Apoptosis Detection Kit (40302ES50, Yeasen Biotechnology, Shanghai, China), following the manufacturer’s instruction. The harvested cells were washed twice with PBS, and were re-suspended in 100 μL 1×binding buffer. Subsequently, the mixture was incubated with 5 μL Annexin V-FITC and 10 μL PI Staining Solution at room temperature for 10-15 min in the dark. Finally, the apoptosis was analyzed by flow cytometry (Bechman, CytoFLEX, USA).