NIK in microglia does not play a role in EAE disease development. (A) Fold change in Map3k14 (NIK) expression relative to the control, calculated using the ΔΔCT method. Expression was measured in isolated microglia (CD11b⁺ cells from the brain) 10 wk after tamoxifen administration. (C) Disease course of NIK^ΔMG and littermate controls. (B) Mice were injected with tamoxifen at 2 wk of age to induce the deletion of NIK in CX3CR1⁺ cells. After 6 wk, mice were immunized with MOG_35–55/CFA and PTx. (D) Maximum EAE score. (E) Representative flow cytometry plots and the number of microglia (CD11b⁺CD45^int), infiltrating CD45^high and CD11b⁺CD45^high immune cells into the CNS (brain and spinal cord) during the peak of EAE (16 dpi). (F) MFI of different activation markers on microglia. (G–I) Percentages of infiltrating CD4⁺CD90⁺ T cells, (H) MOG-responding CD40L⁺CD44⁺ T cells, and (I) cytokine-producing T cells into the CNS, after being subjected to a MOG_35–55 antigen recall assay. Data in graphs are shown as the mean ± SEM and analyzed using two-tailed unpaired Student’s t test or two-way ANOVA with Šídák’s multiple comparisons test. ***P < 0.001. dpi = days after immunization. Each dot represents one mouse. Data are from three independent experiments. MFI, median fluorescence intensity.