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Figure S1. Morphometric evaluation of immunohistological changes in the spinal cord in demyelinating Theiler's murine encephalomyelitis (TME). Box-and-whisker plots show the median and quartiles of the percentage of (A) myelin basic protein (MBP)-positive white matter area, (B) the density of nerve/glial antigen 2 (NG2)-positive cells, (C) glial fibrillary acidic protein (GFAP)-positive cells, and (D) TMEV-antigen positive cells in the lesions and normal appearing white matter (NAWM) of TME virus (TMEV)- and mock-infected SJL/JHanHsd mice, respectively. Significant differences between the TMEV- and mock-infected groups as revealed by Mann–Whitney U-tests, and between normal appearing white matter (NAWM) and lesioned area within the TMEV-infected mice as detected by Wilcoxon signed rank tests are marked as follows: *P ≤ 0.05.

Figure S2. Leading edge analysis of the 26 Gene Ontology (GO) terms significantly co-regulated with the density of nerve/glial antigen 2-positive oligodendroglial progenitor cells in the spinal cord in chronic demyelinating Theiler's murine encephalomyelitis as revealed by Gene Set Enrichment Analysis. The gene set to gene set graph points out that some of these GO terms display major overlap of the comprised genes. Colour intensities show the overlap between subsets: the darker the colour, the greater the overlap between the subsets.

Figure S3. Rotarod performance in Theiler's murine encephalomyelitis virus (TMEV)- and mock-infected SJL/JCrl mice. Box-and-whisker plots show the median and quartiles of the rod speed at the time of drop in revolutions per minute (rpm). The TMEV-infected mice display a chronic progressive decrease in motor performance in the demyelinating phase of the disease. Significant differences between the groups as detected by Mann–Whitney U-test are marked as follows: *P ≤ 0.05.

Figure S4. Histopathology of chronic multifocal to coalescing lymphohistioplasmacytic leucomyelitis with demyelination in Theiler's murine encephalomyelitis. (A) A representative transversal section from the thoracic spinal cord of a TMEV-infected SJL/JCrl mouse at 147 days post infection displays severe coalescing meningeal and perivascular infiltrates consisting of lymphocytes, macrophages and plasma cells (arrow) as well as moderate intraparenchymal infiltration of microglia/macrophages and lymphocytes. Haematoxylin and eosin. (B) A representative transversal section from the thoracic spinal cord of a TMEV-infected SJL/JCrl mouse at 245 days post infection displays extensive demyelination characterized by a loss of blue stain in the white matter (asterix). Luxol fast blue-cresyl violet. (A, B) Scale bars = 100 μm.

Figure S5. Phenotypic characterization of BO-1 cells cultured in (AF) B104-conditioned Dulbecco's modified Eagle's medium (DMEM proliferation medium, (GI) B104-conditioned DMEM with 5 μM retinoic acid (oligodendrocytic differentiation medium, and (JL) DMEM with 20% foetal calf serum (astrocytic differentiation medium. Nuclei were detected using bisbenzimide fluorescence (blue in A, D, G, J, C, F, I, L). Immunofluorescence was performed using a Cy2-coupled anti-bromodeoxyuridine (BrdU)-antibody (green in B, C), and antibodies specific for A2B5 (red in E, F), 2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNPase, red in H, I), and glial fibrillary acidic protein (GFAP, red in K, L) detected by Cy3-conjugated secondary goat anti-mouse antibodies. (E, F) The A2B5-positive BO-1 cells exhibit a bi- or oligopolar phenotype resembling primitive glial progenitor cells. (H, I) The CNPase-positive BO-1 cells exhibit a multiprocessed phenotype resembling immature oligodendrocytes. (K, L) The GFAP-positive BO-1 cells exhibit a hypertrophied, oligopolar phenotype resembling reactive astrocytes. (AL) Scale bars = 100 μm.

Figure S6. Quantitative assessment of the basal proliferation rate, necrosis, apoptosis, STAT3 activation, and phenotypic differentiation of BO-1 cells cultured in B104-conditioned Dulbecco's modified Eagle's medium (DMEM, proliferation medium, B104), B104-conditioned DMEM with 5 μM retinoic acid (oligodendrocytic differentiation medium, B104 RA), and DMEM with 20% foetal calf serum (astrocytic differentiation medium, DMEM/20%FCS). Bar graphs show the mean percentage and standard deviation of three independent experiments for (A) cell proliferation using BrdU labelling, (B) ethidium homodimer III (EthD-III)-positive necrotic cells, (C) FITC-annexin V-positive apoptotic cells, (D) p-STAT3-positive nuclei, (E) A2B5-positive cells, (F) 2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNPase)-positive cells, and (G) glial fibrillary acidic protein (GFAP)-positive cells. Significant differences between the groups as detected by Fisher's exact tests are marked as follows: *P ≤ 0.05.

Figure S7. Quantitative assessment of the basal proliferation rate and phenotypic differentiation of primary rat oligodendroglial progenitor cells cultured in B104-conditioned Dulbecco's modified Eagle's medium (DMEM, proliferation medium, B104), B104-conditioned DMEM with 5 μM retinoic acid (oligodendrocytic differentiation medium, B104 RA), and DMEM with 20% foetal calf serum (astrocytic differentiation medium, DMEM/20%FCS). Bar graphs show the mean percentage and standard deviation of four independent experiments for (A) 2′, 3′-cyclic nucleotide 3′-phosphodiesterase (CNPase)-positive cells, and (B) glial fibrillary acidic protein (GFAP)-positive cells. Significant differences between the groups as detected by Fisher's exact tests are marked as follows: *P ≤ 0.05.

Table S1. Manually curated list of 229 genes involved in Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signalling. The table lists the probe set IDs, gene symbols, as well as the fold changes and P-values obtained at 14, 42, 98 and 196 days post infection. Bold numbers indicate a significant difference between TMEV- and Mock-infected SJL/JHanHsd as detected by Mann–Whitney U-test (P ≤ 0.05).

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