The detection of macrophage surface antigen molecule was used flow cytometry. RAW264.7 cells were plated into a 12-well plate at 2 × 104 cells/well for 24 h, and then were treated with CLDENs, CrDDENs, CrCDENs or LPS. After 24 h of incubation, cells were suspended and incubated with FITC-anti-CD86 antibody (Biolegend, CA, USA), APC-anti-CD206 antibody (Biolegend, CA, USA), PE-anti-MHC II antibody (Invitrogen, Carlsbad, USA) at room temperature for 15 min, and then tested by flow cytometer.
Detection of FITC-dextran internalization was performed to investigate the phagocytosis of RAW264.7 cells. RAW264.7 cells exposed to different concentrations of CLDENs were collected. After incubated with FITC-dextran (1 mg/ml, Angfeibio, Guangzhou, CHN) at 37 °C for 1 h, RAW264.7 cells were treated with cold PBS to terminate the reaction. Then, cells were washed and re-suspended in PBS. The uptake of FITC-dextran in RAW264.7 cells was detected by flow cytometry.
Detection of FITC-dextran internalization was performed to investigate the phagocytosis of RAW264.7 cells. RAW264.7 cells exposed to different concentrations of CLDENs were collected. After incubated with FITC-dextran (1 mg/ml, Angfeibio, Guangzhou, CHN) at 37 °C for 1 h, RAW264.7 cells were treated with cold PBS to terminate the reaction. Then, cells were washed and re-suspended in PBS. The uptake of FITC-dextran in RAW264.7 cells was detected by flow cytometry.