Annexin 5 isothiocyanate

The apoptosis rate was assessed by flow cytometric analysis. The harvested GBM cells were washed twice with cold phosphate‐buffered saline (PBS). Then, these cells were fixed with cold 70% ethanol overnight and stained with 5 μL annexin V‐isothiocyanate (FITC) and 5 μL PI (Keygentec, Nanjing, Jiangsu, China) for 5–15 min in the dark at room temperature. The cells were then examined by flow cytometry (BD Biosciences, Franklin Lakes, NJ).

Briefly, after culturing, treated HepG2 cells were harvested and washed twice with phosphate buffered saline (PBS). Next, the cells were fixed in 70% ethanol for 2 h and then incubated with annexin V-isothiocyanate (FITC) and propidium iodide (PI) (Keygentec, Nanjing, Jiangsu, China) for 10 min in the dark. Finally, flow cytometry (BD Biosciences, Franklin Lakes, NJ, USA) was used to analyze apoptosis. For cell cycle, cell cycle analysis kit (with RNase, Bioss, Cat. No. BA00205) was applied according to experimental instructions. After incubation for 15 min in darkness, apoptosis and cell cycle were examined by flow cytometry and the data were calculated with the CellQuest™ software (648089, Becton-Dickinson, USA).