2.5 Apoptosis detection by flow cytometry
An Annexin V-PE/7-AAD Apoptosis Detection Kit (Yeasen, China) was used to detect apoptosis. Briefly, RAW264.7 cells at 2×105/well were inoculated into six-well plates. After treatment, cells were collected, washed in pre-cooled flow cytometry staining buffer, labeled with Annexin V-PE for 30 min at 4 °C in a dark room, incubated with 7-AAD, and analyzed using flow cytometry (Agilent Technologies, China) and FlowJo software (BD Biosciences, USA).