Flow cytometry

The AsPC-1 and BxPC-3 cells were collected and pretreated with 70% ethanol for 1 h on ice. Afterward, the cells were washed by PBS and incubated with appropriate RNase for 30 min at 37°C. For cell cycle, the cells were labeled using propidium iodide (PI), and the cycle distribution of PDAC cells was analyzed by a flow cytometer. For cell apoptosis, PDAC cells were dual stained using PI and Annexin-V-fluorescein isothiocyanate (FITC) in the dark, and then the percentages of cell apoptosis were determined by the FACSCanto II (BD Biosciences). Each sample was conducted in triplicate and repeated three times.