Flow cytometry (FCM) analysis

The treated RAW264.7 cells were washed once in pre-cooled PBS and then sealed with 100 μl 0.2% bovine serum albumin in PBS for 45 min. After PBS washing, the RAW264.7 cells were incubated with FITC-labeled anti-CD86 (105005, BioLegend, San Diego, CA, USA) in dark at 4°C for 45 min, followed by the detection of the percentage of CD86-positive (CD86⁺) cells on FACSCalibur via Cell-Quest software (Becton-Dickinson, San Jose, CA, USA).