Supplementary MaterialsAdditional file 1. ANOVA with Tukey-Kramer post hoc test. ns, not significant; *p 0.05; ***p 0.001. e H&E staining of longitudinal sections of hind limb muscle of IgG(con) mice, and IgG(AQP4+) mice treated with vehicle or Mem 60 at 8 dpi. Isoalantolactone Scale bars = 100 m. 12974_2020_1913_MOESM1_ESM.tif (1.1M) GUID:?F1DEC7C0-0EF6-4B8F-BE3D-9BD16AA66660 Additional file 2. Infiltration of human IgG in different animal groups. a Representative photomicrographs of cross sections showing the white matter in ventrolateral region of cervical spinal cord. Immunofluorescence staining of human IgG in the spinal cords of IgG(AQP4+) mice treated with preventive memantine or vehicle at 1, 4, 8 dpi; or with therapeutic memantine or vehicle at 8 dpi. IgG(con) mice treated with vehicle were used as a sham control. Sections were counterstained with DAPI. b Lower magnification photomicrographs of spinal cord cross sections showing human IgG immunofluorescence. c-d Representative photomicrographs of brain stem (area postrema) cross sections c and optic nerve longitudinal sections d showing human IgG immunofluorescence. Scale bars = 100 m. 12974_2020_1913_MOESM2_ESM.tif (1.5M) GUID:?32B0C5FD-D133-4548-8B5D-400C2957D484 Additional file 3. Histopathology of spinal cords Itga1 of mice in different groups. Pictures are representative photomicrographs of cross sections showing the white matter in ventrolateral region of cervical spinal cord. Luxol fast blue and H&E staining of the spinal cords of IgG(AQP4+) mice treated with preventive memantine or vehicle at 1, 4, 8 dpi; or with restorative memantine or automobile at 8 dpi. IgG(con) mice treated with automobile had been used like a sham control. Size pub = 100 m. 12974_2020_1913_MOESM3_ESM.tif (2.6M) GUID:?778E2A56-28FA-4755-8E31-B49F6B814C67 Extra document 4. No T cells infiltration towards the spinal-cord. a Immunofluorescence staining of Compact disc4 (T helper cell Isoalantolactone marker) and Compact disc8 (T cytotoxic cell marker) in the vertebral cords of IgG(AQP4+) mice treated with restorative memantine or automobile at 8 dpi. IgG(con) mice treated with automobile had been used like a sham control. Spleens had been used like a positive staining control. Areas had been counterstained with DAPI. Photomicrographs are reps of 5 pets from each combined group. Size bar = 100 m. 12974_2020_1913_MOESM4_ESM.tif (646K) GUID:?27B2B6A6-F613-4F36-9958-F7B47B056FEB Additional file 5. Memantine decreases AQP4 and GFAP loss in the area postrema and optic nerves of IgG(AQP4+) mice. a Representative photomicrographs of brain stem cross sections showing the area postrema. Double immunofluorescence staining of AQP4 and GFAP in IgG(AQP4+) mice treated with therapeutic memantine or vehicle at 8 dpi. IgG(con) mice treated with vehicle were used as a sham control. b Relative intensity of AQP4 and GFAP immunofluorescence in the area postrema. c Representative photomicrographs of longitudinal sections of the optic nerve region proximal to the optic chiasm. Double immunofluorescence staining of AQP4 and GFAP in IgG(AQP4+) mice treated with therapeutic memantine or vehicle at 8 dpi. IgG(con) mice treated with vehicle were used as a sham control. d Relative intensity of AQP4 and GFAP immunofluorescence in the optic nerve. Dotted lines demarcate the areas where quantifications of fluorescence intensities were performed. n = 3 per group. Data are mean SEM. One-way ANOVA with Tukey-Kramer post hoc test. ns, not significant; ***p 0.001. Scale bar = 100 m. 12974_2020_1913_MOESM5_ESM.tif (2.0M) GUID:?C8088E5B-8294-4339-85A6-CAF0224F795B Additional file 6. Memantine reduces demyelination and axonal loss in the area postrema and optic nerves of IgG(AQP4+) mice. a Representative photomicrographs of brain stem cross sections showing the area postrema. Double immunofluorescence staining of MBP and NF-H in IgG(AQP4+) mice treated with therapeutic memantine or vehicle at 8 dpi. IgG(con) mice treated Isoalantolactone with vehicle were used as a sham control. b Relative intensity of MBP and NF-H immunofluorescence in the area postrema. c Representative photomicrographs of longitudinal sections of the optic nerve region proximal to the optic chiasm. Double immunofluorescence staining of MBP and NF-H in IgG(AQP4+) mice treated with therapeutic memantine or vehicle at 8 dpi. IgG(con) mice treated with vehicle were used as a sham control. d Relative intensity of MBP and NF-H immunofluorescence in the optic nerve. n = 3 per group. Dotted lines demarcate the areas where quantifications of fluorescence intensities were performed..