Fig. 2

WML distribution in early and late MOG-EAE and differential axonal injury in early and late chronic MOG-EAE. A Cartoon depicting the analytical strategy of SC sections and anatomical classification of WML, with (w.) and without (w.o.) CSF contact (cont.). B Left: representative SC section stained with luxol fast blue (LFB) and H&E. Scale bar: 500 µm. Black box indicates an anterolateral WML with contact to CSF or pia. Right: CD4 staining in WML and NAWM. C Mean total WML sizes in percent of total white matter area per section from early and late chronic MOG-EAE (n = 8 per condition, two-tailed Student’s t-test). D–F Heat maps depicting the distribution of WML sizes in early and late MOG-EAE with respect to D the anatomical location (cervical, thoracic or lumbar); E the anatomical WML location in anterolateral or dorsal areas; and F the anatomical WML location and contact to the CSF or pia. One-way ANOVA with post hoc Dunnet ‘s test (n = 8 per condition). G Heat map depicting the association of sNfL levels with anatomical WML locations in early and late MOG-EAE (n = 8 per condition, Spearman’s rank correlation). H–I Correlation of sNfL levels with lumbar anterolateral WML locations in H early and I late chronic MOG-EAE (n = 8 per condition, Spearman’s rank correlation). *p < 0.05, **p < 0.01. J Representative images of SC WML and NAWM of MOG-EAE mice in early and late EAE. SMI32-positive axons (arrows) indicate damaged axons. DAPI staining indicates the inflammatory infiltrate. 40 × magnification. Scale bar: 50 µm. K Quantification of mean SMI32-positive axons and WML/NAWM per 200 × 200 µm region of interest (ROI) in early and late chronic EAE (n = 8 mice per condition). Two-tailed Student’s t-test and Mann–Whitney U. Mean and SEM. L, M Association of sNfL levels with axon WML/NAWM ratios in F early chronic and G late chronic MOG-EAE. Pearson’s rank correlation (n = 8 per group)