Annexin 5 pe 7 aad apoptosis detection kit

The percentage of apoptotic cells was analyzed using the Annexin V PE/7-AAD Apoptosis Detection Kit (Beyotime, China).^{43 (link)} A549 and H157 cells were inoculated at an appropriate density on 6-well plates (3 × 10⁵ cells/well) and transfected at 50%–70% confluence. After 48 h, cells were collected and suspended in 1×Binding Buffer (500 μL, P-CA-151, Pricella, Wuhan, China). Cell suspensions were mixed with Annexin V/PE (5 μL, room temperature) and 7-AAD (5 μL, room temperature) for 20 min in the dark. After the completion of the reaction, the stained positive samples were evaluated using flow cytometry immediately (Bio-Rad, USA). FSC-A and SSC-A were used to select the target cell populations. FSC-A and FSC-H were used to remove the adhesive cells. Fluorescein isothiocyanate (FITC) and Percp were used to evaluate apoptosis.