Cell apoptosis and death analysis
Flow
cytometry (Beckman Coulter, Fullerton, California, USA) is used to
distinguish the apoptotic period and quantity through different periods
of apoptosis and different membrane permeability to dyes. In order to
detect the apoptosis rate of the drug, the A549 cell density was
adjusted to 2 × 105 cells / well in a 6-well plate, and the next day was
exposed to final concentrations of 0, 5, 10, 20, and 40 μM NCTD.
According to the manufacturer’s protocol, Annexin V-FITC / PI (Becton
Dickinson, USA) was used to perform the operation. The translocation of
phosphatidy lserine to the cell surface is an indicator of early
apoptotic cells. Therefore, Annexin V-positive and PI-negative cells
were identified as apoptotic. Cell Quest software (FCM, Becton
Dickinson, USA) was used to determine the apoptotic rate.