Figure 2: Adaptive immunity and antigen presentation are required for fibrinogen-induced demyelination.

(a) Affymetrix microarray gene expression analysis and enrichment of gene ontology (GO) of fibrinogen-injected or ACSF-injected corpus callosum at 12 h post injection. Heatmaps of the 142 genes with ≥1.5 × change in expression between ACSF and fibrinogen. (b) Affymetrix microarray gene expression analysis of fibrin-stimulated rat primary microglia at 6 h in vitro. Heatmap and GO analysis of gene expression profiles of 1,342 genes with ≥1.5 × change in expression between unstimulated and fibrin treatment. (c) Affymetrix microarray gene expression analysis of fibrin-stimulated mouse APCs at 6 h in vitro. Heatmap and GO analysis of gene expression profiles of key genes with ≥1.5 × change in expression (left). Heatmap of the M1-related genes with ≥2.0 × change in expression after fibrin stimulation (right). (d) FACS analysis of CD86 expression in APCs after fibrin stimulation. LPS was used as positive control. Anti-CD11b antibody treatment reduces CD86 expression in APCs. Real-time PCR analysis of CD86 gene expression in BMDMs after fibrin stimulation treated with anti-CD11b or IgG isotype control antibody. Data are presented as mean±s.e.m. (n=4 independent experiments; right). **P<0.01, ***P<0.001, ****P<0.0001 (one-way ANOVA). (e) Demyelination (LFB/PAS) in the corpus callosum of WT, MHC II^−/− or RAG2^−/−γc^−/− mice 7 days after fibrinogen injection. Representative images are shown. Scale bar, 100 μm. Data are presented as mean±s.e.m. (n=6 mice per group). **P<0.01, ***P<0.001 (one-way ANOVA and Bonferroni’s multiple comparisons test). ANOVA, analysis of variance; FACS, fluorescence-activated cell sorting; LFB, Luxol fast blue.