Upregulation of Wnt activity in the hippocampus and the spinal cord correlates with disease severity. (a) Iba1 immunostaining (red) of spinal cord sections from Axin2lacZ/+ EAE mice at day 20. The white dashed line highlights lesion areas; white matter tissue is outlined by yellow dashed line. Nuclei were counterstained with Hoechst (blue). Scale bar, 100 μm. (b) Histogram represents the average number of lesions per section of each EAE time point as mean + SEM. Spinal cord: day 20 (n = 4; 15–35 sections/mouse), day 30 (n = 3; 10–23 sections/mouse), day 50 (n = 3; 10–17 sections/mouse). (c) Representative image of lesion site in the DG. Iba1 (red) immunostaining in hippocampal section from EAE mouse at day 30. Nuclei (grey). Scale bar, 50 μm. (d) Representative image of neural stem cells located in the SGZ. SOX2 (green) and GFAP (red). Arrowheads indicate SOX2+/GFAP+ radial glia-like cells and arrows indicate SOX2+/GFAP+ horizontal progenitors. Nuclei (grey). Scale bar, 25 μm. (e+f) β-gal assay of cortical and cerebellar EAE tissue revealed no changes in Wnt activity. Histograms represent mean + SEM of fold-changes of β-gal activity in EAE mice relative to controls (CFA) set as 1. Day 20 (EAE, n = 6; CFA, n = 3); day 30 (EAE, n = 4; CFA, n = 3); day 50 (EAE, n = 8; CFA, n = 4). (g) qPCR analysis of Axin2 in different CNS parts in acute EAE (day 20). Data represent mean of fold-changes + SEM of gene expression in EAE mice (n = 6) relative to controls (CFA; n = 6) set as 1. Expression of Axin2 was normalized to Gapdh. (d) Regression analysis shows correlation between β-gal activity in inflamed CNS tissues and clinical disease score in EAE animals. Pearson correlation, r = 0.58; p = 0.018 (hippocampus) and r = 0.52, p = 0.036 (spinal cord). Statistics: Two-tailed, unpaired Student’s t-test,* * p