Figure 1

IFNβ is predominantly produced by microglia during the effector phase of EAE. a EAE was induced in C57BL/6 N mice by immunization with 200 μg MOG_35–55 peptide. Pertussis toxin was applied i.p. on d0 and d2. Spleen and spinal cord were isolated at indicated time points after immunization. Relative mRNA expression levels for IFNβ (left) and Isg56 (right) were determined by qRT-PCR. Data are pooled from two independent experiments. n = 6–7. b On d17 after EAE induction mononuclear cells from the spinal cord of C57BL/6 N mice were sorted for CD45 and CD11b expression. Relative mRNA expression of IFNβ was determined by qRT-PCR in the indicated cell populations. c Primary adult microglia cultures were generated from WT and IFNβ^mob/mob mice and stimulated with poly (I:C) for 24 h. IFNβ/YFP expression was analyzed by flow cytometry. d and e Immunofluorescent stainings of primary adult microglia treated as in (c). Cells were stained with anti-Iba1 (d) or anti-TLR3 (e) and for YFP for fluorescent microscopy. Scale bar represents 50 μm. f Primary adult microglia from IFNβ^mob/mob mice were stimulated as in (c) and sorted for CD45⁺ CD11b⁺ IFNβ/YFP⁺ vs. CD45⁺ CD11b⁺ IFNβ/YFP⁻ expression. The relative mRNA expression of RIG-I and MDA-5 was determined by qRT-PCR. g On d17 after immunization the phenotype of IFNβ/YFP expressing cells from the spinal cord of WT and IFNβ^mob/mob mice was determined for CD45 and CD11b by flow cytometry. h Quantification of IFNβ/YFP⁺ cells isolated at d17 after immunization from the spinal cord of IFNβ^mob/mob mice. Shown is one representative experiment out of 3 independent experiments. i Representative dot blot shows an overlay of CD45 and CD11b expression of IFNβ/YFP⁺ (red) and IFNβ/YFP⁻ (grey) cells in the spinal cord of IFNβ^mob/mob mice on d17 after immunization. Error bars represent SEM.