Cell Cycle and Apoptosis Analysis Using Flow Cytometry

According to the manufacturer’s instructions, a cell cycle kit (Beckman Coulter) was used to assess the cell cycle in cultured cells using flow cytometry. While employing the recommended procedures, flow cytometry analysis (Beckman Coulter, Inc., Brea, CA, USA) was utilized to identify cell apoptosis in cultured cells using the Annexin V-FITC kit (BioVision, Milpitas, CA, USA). In brief, cells for all groups were cultured at 5 × 10⁵ cells/T75 flask and incubated overnight. After treatment with taxol (13.0 µg/mL) and CMC-AgNPs (7.9 µg/mL) or medium for 48 h, cells were allowed to grow in a 25 cm³ flask until they achieved 70–80% confluence. The cells were then rinsed in PBS and suspended at 5 × 10³–5 × 10⁶ cells/mL in 1 × binding buffer. Then, we added 100 μL of cell suspensions, 5 μL of dissolved PI, and 5 μL of annexin V-FITC solution, and incubated for 15 min in the dark. Following that, we added 400 μL of ice-cold 1 binding buffer and carefully mixed it. Flow cytometric analysis on a COULTER Flow Cytometer (Beckman Coulter) was used to identify apoptotic cells [54 (link),55 (link)].