Pi apoptosis detection kit

To evaluate the protective effect of aqueous leaf extract of M. oleifera on PEDV induced apoptosis, Vero cells were infected with PEDV in the presence of different concentrations of aqueous leaf extract of M. oleifera for 36 h. Equal PBS was treated as the mock control. The cellular apoptosis of different treated groups was measured by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (A211-01; Vazyme, Nanjing, China). Briefly, the cells were harvested and resuspended in 100 μL binding buffer, then labeled with Annexin V and PI for 10 min. Finally, the stained cells were measured with flow cytometry (BD FACSVerse™, Becton Dickinson, Franklin Lakes, NJ, USA), and the data were analyzed using FlowJo software (Version 10, Ashland, OR, USA).

The apoptosis capacity of A172 and U251 cells was tested with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (Vazyme) in line with the manufacturers’ instructions. The results were evaluated with flow cytometry (Beckman Coulter, Atlanta, GA, USA).

The level of cell apoptosis was assessed by flow cytometry with an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit (Vazyme, A211, Nanjing, China) according to the manufacturer’s protocol. In brief, cells were seeded in a six-well plate and transfected with a negative control or siDRD2 or overexpression plasmids and then incubated for 48 h. Afterwards, cultured cells were harvested by trypsinization and cells were washed twice with PBS and resuspended in Annexin V-binding buffer. Cell suspension was then incubated with 5 μL of Annexin V (AV)-FITC and 5 μL of PI staining solution in the dark for 10 min, and the cells were read by a FACS can flow cytometer. A total of 10,000 cells were detected, and to calculate the apoptosis rate, FlowJo v7.6 software (Stanford University, Stanford, CA, USA) was used to analyze the data.