Fig. 7. Improved clinical EAE score following difluorosamine treatment in NG2^CreER:MAPT^mGFP mice is associated with the altered balance of immune cell subsets.

a NG2^CreER:MAPT^mGFP mice were used where GFP expression denotes newly formed oligodendrocytes and myelin. b Representative images of new myelinating oligodendrocytes stained with GFP (green) and olig2 (red). c, d Representative images of longitudinal sections comparing control and peak (Day 18 post-inoculation) or post peak (Day 40) clinical severity of EAE. Stains were GFP (green), MBP for myelin sheaths (yellow) and CD45 (red). Dotted white lines indicate the lesion areas magnified in d. Similar results were noted in a separate experiment. e NG2^CreER:MAPT^mGFP mice were injected with difluorosamine (DIF, 25 mg/kg) starting three days post clinical onset of EAE, for 15 days (treatment period shown by the green box), and average EAE daily clinical score (mean  ±  SEM) is shown. n = 8 mice in each group from 2 independent experiments of 4 mice. Two-way repeated-measures ANOVA with Sidak’s post-hoc test: *p = 0.03, ***p  <  0.001. f–h Spinal cords following treatment were immunostained for CD45, IBA1 and CD4. i–k Bar graphs (mean ± SEM) of number of CD45⁺, IBA1⁺ or CD4⁺ cells in the white matter or lesion ROI. Data are from one representative experiment of 4 mice in each group, where each dot represents one mouse where a mean of 10 lesions were analyzed per mouse. One-tailed unpaired Student’s t test: *p = 0.04. This data was reproduced in a second experiment (n = 4 mice). l–s mRNA expression (mean ± SEM) analyses of lumbar spinal cords comparing the levels of IL-17, IL-10, IL-17/IL-10 ratio, IL-12, arginase-1 (Arg1), inducible nitric oxide synthase (iNOS) and IL-1α between DIF and vehicle treated EAE mice. Data pooled from two independent experiments, n = 8 mice, two-tailed unpaired Student’s t test; *p = 0.01, ***p < 0.001. Source data are provided in the source data file.