(A) Foxo3−/− (open circles, bars) and WT littermate mice (black circles, bars) were immunized with 50μg of peptide MOG35–55 emulsified in CFA at day 0 and 200 ng of pertussis toxin was injected iv. on day 0 and day 2 (n=14 mice per genotype) (B) 2D2-Foxo3−/− (open circles, bars) or 2D2-WT (black circles, bars) were injected iv. with 150 ng of Pertussis Toxin at day 0. (n=6 mice per genotype) (C) Rag2−/−-Foxo3−/− (open circles, bars) or Rag2−/−-Foxo3+/+ (black circles, bars) mice were injected iv. with 2.104 2D2-WT naive CD4+ T cells mixed with 4.106 WT CD4+ T cells. Mice were then immunized with 50μg of peptide MOG35–55 emulsified in CFA and injected iv. with 100 ng of pertussis toxin. (n=6–7 mice per genotype). (D) Foxo3fl/fl-Cd4-cre+ (open circles, bars) or Foxo3fl/fl-Cd4-cre− (black circles, bars) littermate controls were immunized as in A. Incidence and mean cumulative clinical scores are shown (n=11–14 per genotype). Incidence, clinical scores and mean with SEM of cumulative clinical scores were calculated. Error bars, s.e.m.; P values (Mann–Whitney U test); P values for clinical Scores (2way ANOVA). Data are representative of at least three independent experiments. See also Figure S6