Clinical score and co-localization of TNFR1 in EAE-induced mouse brain. EAE mouse model was immunized with MOG and CFA, and scored daily as described in "Methods". Mice received anti-CD40 antibody (1 mg/kg), 8-oxo-dG (60 mg/kg) or a combination of anti-CD 40 antibody and 8-oxo-dG, respectively, with i.p. injection for 5 days after MOG injection. After animal sacrifice at day 32 (EAE score, 3.8 ± 0.21), brains were removed and preserved in 10% neutral buffered formalin. Expressions of TNFR1 were determined in protein extracts by western blot, and co-localization was determined by double staining for TNFR1 (green) and GFAP (red). (A) EAE score after pretreatment with inhibitors Data are mean ± SEM (n = 8). (B) Expression of TNFR1 (n = 4). Histogram for densitometry analysis was indicated by mean ± SEM (n = 4) obtained from four independent experiments. (C) Co-localization of TNFR1 and astrocytes after pretreatment with inhibitors (n = 4). The image depicted one of representative results after four independent experiments were determined. TNFR1, TNF-α receptor 1; αCD40, anti-CD40 antibody; 8-oxo-dG, 8-hydroxydexoyquanosine; both, a combination of anti-CD40 antibody and 8-oxo-dG. Bar in control group indicates 100 μm.