2.5. Flow cytometry assay

Glioma cells were plated into 6‐well plates, and the Annexin V‐FITC apoptosis assay kit (BD Biosciences, USA) was implemented to evaluate apoptosis of cells in different treatment groups. At room temperature, cells were digested by trypsin and washed. Afterward, the FACSCanto Ⅱ flow cytometry instrument (BD Biosciences) was applied for cell apoptosis analysis, with the FITC‐labeled Annexin V as the fluorescence probe. The assay was repeated three times.

FACSCanto Ⅱ flow cytometry instrument (BD Biosciences, USA) was introduced for cell cycle assay at 48 h after mimic transfection. Note that 1 × 10⁶ cells were collected, followed by washing with cold PBS, and fixing with 70% cold ethanol at 4°C for 48 h. Next, after rinsing cells with cold PBS again, propidium iodide (PI) was used for staining nucleus followed by measuring DNA amount using flow cytometry. At least 1 × 10⁴ cells were counted, and the cell cycle distribution was analyzed using ModFit software.