Flow cytometry assay
Single suspensions of mouse mesenteric lymph nodes or peripheral blood mononuclear cells (PBMCs) were prepared. Then the single cells were blocked with purified anti-mouse CD16/CD32 mAb for blocking and flow cytometry antibodies: BUV395-conjugated CD11b, FITC-conjugated CD11c, PE-conjugated Ly6G, BV510-conjugated F4/80, APC-conjugated Ly6C, BUV395-conjugated CD4, Percy-cy5.5-conjugated CD3 and PE-conjugated CD69. After incubation at 4℃ for 20 min, the cells were washed once with phosphate buffer saline and analyzed using a BD LSR Fortessa Flow Cytometer. The data were analyzed with FlowJo software.