Extended Data Fig. 3. Gating strategy for the characterization of AQP4-specific T cell phenotypes.

(a) Representative cytograms of stainings for transcription factors Foxp3, ROR-γt, T-bet, and Bcl-6 in all vs. AQP4-specific CD4⁺ T cells isolated from the spleens of P41-immunized Aqp4^–/– mice. A dump channel contained the markers CD11b, CD19, NK1.1, and F4/80. (b,c) Chimerism of the B cell compartment in mixed bone marrow chimeras harbouring AQP4-sufficient or AQP4-deficient B cells. Bone marrow cells from B cell-deficient (ΔB) homozygous Mb1-Cre^KI/KI x Aqp4^–/– mice (CD45.2) and either AQP4-competent wild-type or AQP4-deficient Aqp4^–/– mice (CD45.1) (9:1) were grafted into Aqp4^–/– mice (CD45.1^+/–) to generate mixed bone marrow chimeras (MBMC), in which only B cells were competent in AQP4 expression in an otherwise AQP4-deficient environment. After six weeks, the lymphoid compartments (SPL, spleen; THY, thymus) were fully reconstituted, with the B cell compartment largely derived from the CD45.1⁺ AQP4-competent or deficient donors, respectively. The diagram in (b) was created using Servier Medical Art under a Creative Commons licence CC BY 3.0.