Fxp0211

Cells were cultured in 6‐well plates for 48 h, the serum‐free medium was replaced and cultured for 12 h. Then, the complete medium was replaced and the cells were collected after being cultured for another 24 h. After washing with pre‐cold phosphate buffered saline (PBS), the cells were incubated with 95% ethanol at 4°C for 48 h. Then, cells were washed with pre‐cold PBS again and stained with PI (FXP0211,4A biotech, China) according to the instructions of the manufacturer. The cell cycle was analysed by the Millipore Guava Flow cytometer (Millipore Corp, Bedford, MA, USA).

Cell cycle analysis was measured by a cell cycle and apoptosis analysis kit (FXP0211, 4A BIOTECH, Beijing, China). Briefly, the cells were fixed and stained with PI. Flow cytometry was performed using the BD Flow Cytometry Analyser. The apoptotic activity in cells was measured by an Annexin V-FITC/PI apoptosis detection kit (FXP018, 4A BIOTECH, Beijing, China), according to the manufacturer’s instructions. Briefly, the cells were washed with pre-cooled PBS and resuspended in Annexin V binding solution. Next, the cell suspension was incubated with Annexin V-FITC in the dark and then with Propidium Iodide. The cells were analyzed by flow cytometry.