Fig. 2

Flow cytometry gating strategy for identification of ILCs, LTi, and T_H17 cells in EAE mice. Forward scatter area (FSC-A) and height (FSC-H) were used to exclude cells, which had formed doublets. The lymphocyte gate was determined by using FSC-A and sideward scatter area (SSC-A). Viable cells were separated from dead cells. Cells were divided into CD3⁻CD5⁻ and CD3⁺CD5⁺ cells. CD3⁻CD5⁻ cells were further analyzed for the presence of LTi cells and ILCs, and CD3⁺CD5⁺ cells were further analyzed for T_H17 cells. CD3⁻CD5⁻ cells being positive for CD4 and RORγt represented CD3⁻CD5⁻CD4⁺RORγt⁺ LTi cells. Moreover, CD3⁻CD5⁻CD4⁻RORγt⁺ cells appeared in the ILC gate. CD3⁺CD5⁺ cells, which were also positive for CD4 and RORγt, occurred in the gate of T_H17 cells