Fig. 3

Prophylactic and semi-therapeutic ibrutinib treatment in Biozzi EAE reduce spinal cord demyelination, inflammation, and white matter cell apoptosis. Analyses were performed on all surviving mice (n = 11–13 mice/group) from the experiment shown in Fig. 2a. The figure legend in d applies to all graphs. a-c Illustrative thoracic spinal cord sections stained with anti-MBP antibody (brown) and hematoxylin (nuclei; blue). Adjustments to gamma were made evenly across these images to better visualize staining. Scale bars, 100 µm. d Mean demyelination scores per section, per mouse. e–g Illustrative thoracic spinal cord sections stained with hematoxylin (nuclei; blue) and eosin (cytoplasm; pink). Scale bars, 100 µm. h Mean inflammation quantified as number of 20+ cell foci per section, per mouse. i–k Magnified white matter regions from e–g demonstrating inflammatory infiltrates. Arrows in i identify apoptotic nuclei, with magnified inset demonstrating an apoptotic nucleus with karyorrhexis. Scale bars, 10 µm. l Mean numbers of apoptotic nuclei quantified per section, per mouse. Data in d, h, and l were obtained by taking the mean value of 3 spinal cord regions (cervical, thoracic, and lumbar) from 1 slide per stain, per mouse. Graphed data are shown as mean ± SEM. Significance was tested using a Kruskal–Wallis test followed by Dunn’s multiple comparisons test (d) or one-way ANOVA followed by Dunnett’s multiple comparisons test (h, l). Asterisks denote results of multiple comparisons tests. *p < 0.050, **p < 0.010