Flow cytometer

For measuring differentiation of mø, the flow cytometer was used. We used anti-CD11b PE/cy7, anti-CD11c PerCP/cy5.5, anti-F4/80 PE, and anti-CD206 allophycocyanin (BioLegend) to stain BMDM. Staining was performed in PBS supplemented with 0.1% BSA and 0.02% sodium azide for 30 min at 4°C. Intracellular staining was performed by the protocol described by the manufacturer (BioLegend).

Cells were examined on the FACS Canto flow cytometer and analyzed by FlowJo v10.0.7 software. mø were defined as CD11b⁺F4/80⁺ cells, M1 mø were defined as CD11b⁺F4/80⁺CD11c⁺CD206⁻ cells, and M2 mø were defined as CD11b⁺F4/80⁺CD11C⁻CD206⁺ cells.