Fig 2: Astrocytes express proliferation markers in chronic EAE. (a) Correlation of module eigengenes with EAE disease progression from unimmunized control to chronic EAE. Numbers indicate Pearson r and p value in brackets. Significant modules are labelled in bold. (b) Mean expression of eigengenes from the blue, yellow, and turquoise modules in different conditions, depicted as row z-scores (top), and selected enriched GO terms associated with each module (bottom). For a complete list of GO terms per module see supplemental file Table S6. (c, d) Representative images of KI67 (green) and GFAP (magenta) co-expression in mouse spinal cord during EAE disease progression (c). Enlarged image shows co-expression in spinal cord of chronic EAE mice (d) (n = 3)

Fig 3: Implanted HAP stem cells inhibit glial scar formation at the ICH site in C57BL/6J.(A, B) Expression of GFAP of astrocytes in the ICH area with and without HAP-stem-cell-implantation in C57BL/6J mice. Glial scar formation was observed in the non-implanted control mice but not HAP-stem-cell-implanted mice. (C, D) Expression of Iba-1 of microglia in the ICH area with and without HAP-stem-cell implantation in C57BL/6J mice. (E) The GFAP-positive area in coronal section of the brain in C57BL/6J mice was smaller in the HAP-stem-cell-implanted mice than in the non-implanted control mice (n = 4, each group). *P < 0.05. (F) The Iba-1-positive area in coronal sections of the brain in C57BL/6J mice was smaller in the HAP-stem-cell-implanted mice than in the non-implanted control mice (n = 4, each group). *P < 0.05. Bar = 50 µm. All images show coronal sections of the brain.

Fig 4: The development of superficial layer of retinal vasculature is independent of Vdr expression.(A) Demonstrates GFAP and Col IV stained retinal vessels prepared from postnatal day 5 (P5) Vdr +/+ and Vdr -/- mice. Please note similar expansion of astrocytes (green, GFAP) and progression of expanding vessels (red, Col IV). Scale bar = 200 µm for x25 and Scale bar = 50 µm for x100 images. (B) The mean number of angiogenic sprouts at the angiogenic fronts were quantified per field (x100) in each retina. (C) Coverage of retinal vasculature relative to total retina area were measured for each retina and is shown as a percentage. (n= 5; each point represents one mouse).

Fig 5: Reactivity of spinal cord astrocytes is most pronounced during acute stage of EAE. (a) Upset diagram depicting the number and overlap of DEGs comparing each EAE stage to unimmunized control. Bars show number of enriched and depleted genes (bottom-left). Overlapping DEGs are illustrated by interconnected dots between groups (bottom-right) and the number of DEGs are depicted in in bar graph (top). (b) Circos plot depicting GO annotations of up- and down-regulated genes per EAE stage compared to astrocytes from unimmunized control mice. (c) Mean log expression of pan-reactive, LPS-reactive, and MCAO-reactive astrocyte markers, and genes involved in lactate metabolism, myelination, blood brain barrier, and cholesterol synthesis. For a list of genes see supplemental file Table S1 (n = 4–6). (d) Normalized expression of selected reactive astrocyte markers Serpina3n, H2-D1, and Lcn2 and of C4b, Cxn43, and Btbd17. Significantly different expression compared to unimmunized control is indicated (#). Boxes show 25th to 75th percentiles and median, and whiskers indicate min/max (n = 4–6). (e) Representative images of MHC-II (green) and GFAP (magenta) co-expression in mouse spinal cord during EAE disease progression. Yellow arrows indicate co-expression; white arrows indicated MHC-II expression in GFAPneg structures (n = 3)