Surface staining was performed in PBS containing LIVE/DEAD™ Fixable Aqua Dead Cell Stain (Thermo Fisher Scientific) for 30 min at 4 °C. For intracellular staining, cells were fixed and permeabilized, and intracellular staining was performed in Permeabilization Reagent (Thermo Fisher Scientific). Antibodies (all from BioLegend, unless otherwise stated) against following antigens were used with the indicated dilutions: CD3 (17A2; 1:200), CD4 (RM4-5; 1:200), CD8 (53-6.7; 1:100) CD11b (M1/70; 1:100), CD11c (N418; 1:100), CD25 (PC61; 1:200), CD45 (I3/2.3; 1:100), CD83 (Michel19; 1:50), CD86 (GL-1; 1:200), CX3CR1 (SA011F11; 1:200), FoxP3 (FJK-16s, eBioscience; 1:100), IFN-γ (XMG1.2; 1:200), IL-17A (TC11-18H10.1; 1:300), I-A/I-E (M5/114.152; 1:200), Ly6C (HK1.4; 1:400), Ly6G (1A8; 1:100), P2RY12 (S16007D; 1:100). Expression was assessed with a FACSCantoII flow cytometer (BD) and analyzed with FlowJo software version 10 (FlowJo, LLC). Cells were pre-gated on CD45+ for further analyses.
