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.