Additional file 1 of IFNAR signaling of neuroectodermal cells is essential for the survival of C57BL/6 mice infected with Theiler’s murine encephalomyelitis virus
Creators: name: Bühler, Melanie
name: Li, Dandan
name: Li, Lin
name: Runft, Sandra
name: Waltl, Inken
name: Pavlou, Andreas
name: Kalinke, Ulrich
name: Ciurkiewicz, Malgorzata
name: Huehn, Jochen
name: Floess, Stefan
name: Beineke, Andreas
name: Baumgärtner, Wolfgang
name: Gerhauser, Ingo
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Neuroscience
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Mental Health
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Plant Biology
subject: Virology

description: Additional file 1: Table S1. Summary of primer pairs used for reverse transcriptase polymerase chain reaction. Table S2. Transcriptional changes in the cerebrum of TMEV- and mock-infected SJL and C57BL/6 mice (fold changes). Fig. S1. RT-qPCR of the cerebrum at 4, 7 and 14 days after TMEV-BeAn infection (dpi). lfna mRNA transcripts were downregulated in TMEV- compared to mock-infected SJL and C57BL/6 mice at 14 dpi, whereas no significant impact of TMEV infection on Ifnb1 mRNA transcripts was found. Irf7, Isg15 and Eif2ak1 mRNA transcripts were increased in TMEV- compared to mock-infected SJL and C57BL/6 mice at 4, 7 and 14 dpi except for Isg15 mRNA transcripts in C57BL/6 mice at 14 dpi. Mock-infected SJL: n=5 (4, 7, 14 dpi); TMEV-infected SJL: n=6 (4,7 dpi), n=5 (14 dpi); mock-infected C57BL/6 mice: n=6 (4, 14 dpi), n=5 (7 dpi); TMEV-infected C57BL/6 mice: n=4 (4 dpi), n=3 (7 dpi), n=5 (14 dpi). The mRNA copy numbers were normalized using a normalization factor calculated from three housekeeping genes (Gapdh, Actb, Hprt). Mann Whitney tests: * p &lt; 0.05; ** p &lt; 0.01. Shown are all data points with means. Each data point represents the mRNA copy number of one mouse. Fig. S2. ISG15, OAS1 and PKR protein expression in the brain of TMEV-infected C57BL/6 mice. Shown is the hippocampus of TMEV-infected C57BL/6 mice at 4 (ISG15 and PKR) and 7 (OAS1) days post infection. Immunohistochemistry using the avidin-biotin-peroxidase complex method with the chromogen 3’3-diaminobenzidine and Mayer’s hematoxylin counterstaining. Fig. S3. Hippocampal cell loss at 4 days after TMEV-BeAn infection. No significant differences were found between TMEV-infected IFNAR-/- (n=6), NesCre+/-IFNARfl/fl (n=10), GFAPCre+/-IFNARfl/fl (n=9), Syn1Cre+/-IFNARfl/fl (n=10) and Cre-/-IFNARfl/fl mice (n=7) as well as Tam-treated Sall1CreER+/-IFNARfl/fl (n=6) and Sall1CreER-/-IFNARfl/fl mice (n=9). Shown are all data points with means. Each data point represents the semiquantitative score of one mouse. Fig. S4. Percentages of perivascular CD3+ T cells, CD45R+ B cells and Iba-1+ macrophages in the cerebrum at 4 days after TMEV-BeAn infection. No significant differences in the percentages of perivascular mononuclear cells were found between TMEV-infected IFNAR-/- (n=6), NesCre+/-IFNARfl/fl (n=8), GFAPCre+/-IFNARfl/fl (n=6), Syn1Cre+/-IFNARfl/fl (n=6) and Cre-/-IFNARfl/fl mice (n=6) as well as Tam-treated Sall1CreER+/-IFNARfl/fl (n=6) and Sall1CreER-/-IFNARfl/fl mice (n=9). Shown are all data points with means. Each data point represents the percentage of immunopositive cells of one mouse. Fig. S5. GFAP+, MAC3+, and TMEM119+ cells in the hippocampus at 4 days after TMEV-BeAn infection. No significant differences in the semiquantitative analysis of GFAP+, MAC3+ and TMEM119+ cells were found between TMEV-infected IFNAR-/-, NesCre+/-IFNARfl/fl, GFAPCre+/-IFNARfl/fl, Syn1Cre+/-IFNARfl/fl and Cre-/-IFNARfl/fl mice (n=5-10). Shown are all data points with means. Each data point represents the semiquantitative score of immunopositive cells of one mouse. Fig. S6. A plaque assay confirms the presence of infectious virus in the brain at 4 days after TMEV-BeAn infection. Infectious virus was found in TMEV-infected IFNAR-/-, NesCre+/-IFNARfl/fl, GFAPCre+/-IFNARfl/fl, Syn1Cre+/-IFNARfl/fl and Cre-/-IFNARfl/fl mice as well as Tam-treated Sall1CreER+/-IFNARfl/fl and Sall1CreER-/-IFNARfl/fl mice. Shown are all data points with means. Each data point represents the plaque-forming units/ml of one mouse (n=3). Fig. S7. Nonparametric Spearman correlation coefficients of RT-qPCR data of the cerebrum at 4 days after TMEV-BeAn infection. Fig. S8. Nonparametric Spearman correlation coefficients of TMEV RNA and cytokine array data of the cerebrum at 4 days after TMEV-BeAn infection. Fig. S9. TMEV antigen and IL-6 and IL-10 protein expression in the brain of a TMEV-infected NesCre+/-IFNARfl/fl mouse. Shown is the hippocampus of a TMEV-infected NesCre+/-IFNARfl/fl mouse at 4 days post infection (serial sections of the animal shown in Fig. 3). TMEV: Note high amount of TMEV antigen in the hippocampus. IL-6: There is a strong IL-6 expression by hippocampal neurons (upper inset), whereas most perivascular mononuclear cells lack an IL-6 immunoreaction (lower inset). IL-10: IL-10 immunoreactivity was only found in few small round and elongated cells (upper inset) as well as medium-sized round cells (arrow; lower inset). Immunohistochemistry using the avidin-biotin-peroxidase complex method with the chromogen 3’3-diaminobenzidine and Mayer’s hematoxylin counterstaining (serial sections).