Annexin pe 7 aad apoptosis detection kit

After washing the cells in PBS and adjusting the cell concentration to 2 × 10⁶/mL, the Annexin PE/7-AAD Apoptosis Detection Kit (Vazyme, China) was employed under the manufacturer’s instructions. 1 × 10⁶ cells were rinsed using PBS and fixed in 75% ethanol for the duration of the cell cycle assay. Cell cycle and apoptosis were measured through Coulter FC500 flow cytometry (Beckman, USA).

K562 and K562/G01 cells were collected 72 h after lentivirus infection. For the apoptosis assay, 1 × 10⁶ cells were washed with cold phosphate buffered saline (PBS) and resuspended in 1 ml of 1× binding buffer. Then, the cells were stained with 5 μl of phycoerythrin (PE) and 5 μl of 7-aminoactinomycin D (7-AAD) successively using Annexin PE/7-AAD Apoptosis Detection Kit (Vazyme, Nanjing, China) according to the manufacturer’s instructions. For cell cycle assay, 1 × 10⁶ cells were washed and resuspended in cold PBS and incubated in ice-cold 70% ethanol for 4 h. Coulter FC500 flow cytometry (Beckman, CA, USA) was applied to detect apoptosis and cell cycle [16 (link)].