Apoptosis assay

An apoptosis assay was performed by flow cytometry with Annexin V‐FITC/propidium iodide (PI) staining. After transfection for 24 h, cells were harvested and seeded at 1 × 10⁵ cells per well in 100 μL culture medium in a 12‐well culture plate. The cells were washed twice with PBS and resuspended in 400 μL Annexin V‐FITC binding buffer. Then, 5 μL Annexin V‐FITC and 10 μL PI were added to stain the cells in accordance with the instructions of the Apoptosis Detection Kit (Qihan Biotech, Shanghai, China). Then, apoptosis was detected on a flow cytometer (BD Biosciences, USA). The experiment was repeated three times.