Mod fit lt software v2

According to the standard method, cell cycle analysis was performed. In short, 5×10⁵ cells with 70% cold ethanol under –20°C overnight. The next day, the fixed cells were centrifuged at 1200 x g for 1 min and washed twice with PBS. After the cells with 200 µL RNase A (1 mg/ml) for 10 min at 37°C in suspension, then add 300 µl propidium iodine (PI, 100 mu l/mL, BioVision, Milpitas, CA, USA) to dyeing the DNA in cells in the dark. After being incubated at room temperature for 20 mins, cells in FAC Scan flow cytometer (Becton Dickinson, Franklin ‘lakes, NJ, USA) in the analysis of cellular DNA content, and through the Mod Fit LT software V2.0 (Becton Dickinson) analyzed the data.

Cell cycle analysis was performed using flow-cytometry. For fixation, 5×10⁵ cells were incubated with 70% ice-cold ethanol at −20°C overnight. The next day, the fixed cells were centrifuged at 1,200 × g for 1 min at 4°C and washed twice with PBS. The cells were resuspended in 200 µl RNaseA (1 mg/ml) for 10 min at 37°C, 300 µl PI (BioVision, Inc.) was added, and the cells were incubated at room temperature for 20 min to stain the DNA. The cells were protected from light during the staining procedure. The cells were analyzed for cellular DNA content using Mod Fit LT software V2.0 (Becton Dickinson and Company) with a FACScan flow cytometer (Becton, Dickinson, and Company).

5 × 10⁵ cells with 70% cold ethanol were cultured overnight under –20 °C. Then, the cells were centrifuged at 1200×g about 1 min and washed twice with PBS. Next, the cells were treated with 200 µL RNase A (1 mg/mL) for 10 min in suspension at 37 °C, and 300 µL PI (50 μg/mL, BioVision, Milpitas, CA) was then added into the cells in the dark. After 20 min of incubation at room temperature, cellular DNA content of the cells was analysed using FAC Scan flow cytometer (Becton Dickinson, Franklin Lakes, NJ) and the data were analysed by the Mod Fit LT software V2.0 (Becton Dickinson, Franklin Lakes, NJ).