AMΦ were cultured for a defined time period, fixed in 4% paraformaldehye/1 × PBS for 15 min. Cells were washed three timers with 1 × PBS and permeabilized using 0.1% Triton X-100 in 1 × PBS, and blocked with 5% BSA for 45 min and sequentially administered primary antibody and secondary antibody (Alexa-488-conjugated donkey anti rabbit secondary antibody). Nuclei were stained with DAPI for immunofluoresence analysis. Stained cells were examined and recorded using EVOSfl fluorescence microscopy.