Fig. S3.

T_MOG cell priming in the secondary lymphoid organs is not changed in the presence of B_MOG cells. T_MOG cell activation was analyzed during the priming phase (days 2–4 p.i.) or briefly before disease onset at day 9 p.i. (A and B) T_MOG cell proliferation. (A) Proliferation of T_MOG cells in the spleen of T-/B-MOG and T-MOG mice at the indicated time points. Data are presented as the percentage of dividing T_MOG cells (CSFE dilution). Mean ± SEM, n = 6–10. (B) Numbers of T_MOG-RFP cells at day 9 p.i. in the indicated organ per gram of tissue or per milliliter of blood. Data are presented as mean ± SEM. n = 4–5. (C and D) Expression of membrane activation markers in T_MOG cells at days (C) 4 or (D) 9 p.i. analyzed by flow cytometry. The percentage of positive T_MOG cells is depicted as mean ± SEM, n = 4. (E) Cytokine production evaluated by quantitative PCR in ex vivo-sorted T_MOG-RFP cells (Left) or by ELISA stimulated in vitro with the indicated amounts of MOG_35–55 (Right). Cytokine production was assessed at the indicated days p.i. from the draining lymph node (first and third rows) or the spleen (second and fourth rows). Data are presented as mean ± SEM and represent four to six animals per group. (F) Expression of genes of the T_FH program. Expression levels of the genes coding for Bcl-6, PD-1, ICOS, CXCR5, and IL-21 were analyzed at day 4 p.i. Data for the spleen or draining lymph nodes are presented as mean ± SEM. n = 4.