2.2. EAE Model

We chose to exclusively use female mice for this study, because C57BL/6J males do not display severe clinical symptoms and reach the maximum clinical score [20]. At 13 weeks of age, we induced the EAE model using the MOG 35-55/CFA EMULSION PTX kit (#EK2110, Hooke Laboratories, Lawrence, MA, USA), as previously described [4]. Briefly, an emulsion containing myelin oligodendrocyte glycoprotein (MOG) 35-55 peptide and complete Freund’s adjuvant was subcutaneously injected into each hind leg (0.1 mL/leg). At 2 and 24 h after injection, pertussis toxin (PTX) (100 ng/mouse) was administered intraperitoneally. The control mice were injected with an emulsion containing complete Freund’s adjuvant without MOG 35-55 peptide and PTX. After the induction of immunization, optic neuritis typically starts at 2 weeks and peaks at 3 weeks [5]. The clinical score for EAE (n = 7) and control (n = 5) mice was obtained at 0, 7, 10, 14, 21, and 28 days post-immunization (dpi), in accordance with the manual of the MOG 35-55/CFA EMULSION PTX kit and based on changes in the motor functions of the tail, legs, and neck. Rapid increases in the clinical score usually occur between 12 and 18 dpi [21].