Fig. 3

Histological assessment of neuroinflammation and tissue damage. A IBA1-positive cell densities in DTH and control animals. The mean density of IBA1-positive cells (microglia/macrophages) was greater in DTH animals compared to controls at all time points, with the peak density noted at day 12 (a). Fold change of IBA1-positive cell density within DTH animals, normalized to the mean density of the controls at each time point (indicated by the horizontal dashed line at 1.0), showing that the greatest fold change occurred at day 12 (b). Representative images of IBA1 staining in day 12 control and in DTH animals at all 3 time points (c). B GFAP-positive cell densities in DTH and control animals. The mean density of GFAP-positive cells (astrocytes) was higher in DTH animals compared to controls at all time points (a). Fold change of GFAP-positive cell density within DTH animals, showing that the highest fold change was at day 12 and decreased with time (b). Representative images of GFAP staining in day 12 control and in DTH animals at the 3 time points (c). C Area of MBP loss within DTH animals. MBP (myelin) loss was the least pronounced at day 12, increased at day 28, and plateaued at day 60 (a). Representative images of MBP staining in day 12 control and in DTH animals at all time points (b). D Area of NfH loss within DTH animals. NfH (axonal) loss in DTH animals was the lowest at day 12, and increased at days 28 and 60 (a). Representative images of NfH staining in day 12 control and in DTH animals at all time points (b). ****p < 0.0001, ***p < 0.001, **p < 0.01, and *p < 0.05 by Sidak post hoc test after two-way ANOVA (A.a and B.a) and by Tukey post hoc test after one-way ANOVA (A.b, B.b, C.a, and D.a). All scale bars = 50 μm. Data presented as mean ± SEM. ANOVA analysis of variance, DTH delayed-type hypersensitivity, GFAP glial fibrillary acidic protein, IBA1 ionized calcium binding adaptor molecule 1, MBP myelin basic protein, NfH neurofilament-heavy chain, SEM standard error of mean