Passive EAE induction by adoptive transfer of MOG_35–55–reactive T cells from DR2b donors into DR2b and DR2bΔR2 recipients. (A) Representative IF staining of cerebellum from DR2b and DR2bΔR2 recipients, analyzed at progressive phase of disease, showing myelin stain (FluoroMyelin, red) and nuclear stain (DAPI, blue). White arrows indicate lesions. Scale bar: 200 μm. Pooled data from 3 independent experiments with n = 20 DR2b mice and n = 18 DR2bΔR2 mice per group. (B) Representative IF staining of cerebellum as in A, showing myelin stain (FluoroMyelin, red) and neurons (NeuN, green). Scale bar: 100 μm. Myelin fluorescence intensity data pooled from 3 independent experiments with n = 9 mice per group. (C) Representative IF staining of cerebellum analyzed at onset, acute phase, and progression of disease, showing myelin stain (FluoroMyelin, red), oligodendrocyte precursor cells (Olig2, green), and nuclear stain (DAPI, blue). Scale bar: 200 μm. Shown are representative data with n = 4–5 mice per group from 4 independent experiments. Dashed circles represent Olig2⁺ cell clusters (≥10 cells); white arrows indicate single Olig2⁺ cell staining. (D) CNS Olig2⁺ cell relative abundance in DR2b and DR2bΔR2 recipients at onset, acute, and progression phase of disease determined by flow cytometry. Shown are fold changes relative to mean cell number in DR2b mice at each time point. Representative data of 2 independent experiments with n = 3 mice per group. (E) Concentration of CXCL12 (CXCR4 ligand) at acute and progression phases of disease, and CCL11, CCL7, and CCL5 (CCR3 ligands) in whole CNS tissue homogenate at progression phase of disease. Pooled data of 3 independent experiments. n = 9–18 mice per group. Statistical significance was determined by Student’s 2-tailed t test with Welch’s correction (B–E) or Mann-Whitney U test (U = 90.5) (A). *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001. Error bars indicate mean ± SD.