2.4. Flow cytometry assay

After transfection with pcDNA3.1 or pcDNA3.1‐Dcf1 for 48 h, glioblastoma cells were digested with trypLE, harvested in 10% FBS in DMEM/F12, washed twice with 2% BSA in PBS, resuspended in 100 μl of binding buffer, combined with 5 μl of annexin V‐FITC and 5 μl of PI according to the protocol of an Annexin V‐FITC/PI Apoptosis Detection Kit, mixed gently and stained at room temperature for 15 min before flow cytometry detection. The cell cycle was detected by flow cytometry after digestion with 10 μg/ml RNase at 37℃ for 30 min and staining with 50 μg/ml PI at 4℃ for 30 min. The percentages of apoptotic cells and cells in each phase of the cell cycle were calculated.