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Table 1.

Viral paralysis scale [27].

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Fig 1.

Behavioral changes of mice at different time.

(A) Body weight change of the mice at different time; (B) VPS scores at different time. Kruskal-Wallis H test was used (****p< 0.0001); (C) Hanging wire test at different time. Kruskal-Wallis H test was applied for statistical analysis (**p = 0.005).

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Fig 2.

CMAP assessment of the sciatic nerve in mice.

(A) Amplitude. Kruskal-Wallis H test was used (*P = 0.0173); (B) End latency. One-way ANOVA was used (P = 0.0903); (C) Conduction velocity. Statistical analysis by one-way ANOVA (****P < 0.0001).

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Fig 3.

Immunofluorescence of mouse sciatic nerve at different time.

One-way ANOVA was used. (A) The sciatic nerve tissue was double stained with MBP + NF-H and labeled with fluorescence. MBP was labeled in red and NF-H in green. Nuclei are stained DAPI (blue); (B) MFI of MBP (**P = 0.0015); (C) MFI of NF-H (P = 0.0666).

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Fig 4.

Ultrastructural of sciatic nerve of mice at different time.

(A) 2000 × Magnification; (B) 5000 × Magnification. Magnified image of the arrowhead in (A). (MF) Myelinated nerve fiber, (NMN) Nonmyelinated nerve, (MS) myelin sheath, (A) axon, (SC) Schwann cells, (M) mitochondria, (RER) Endoplasmic reticulum structure.

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Fig 5.

Western blot analysis of related protein expression levels.

Statistical analysis by one-way ANOVA. (A) The protein expression was detected by Western blot. Using GAPDH as the reference, the changes of relative expression of different proteins at different time were compared. (B) Relative expression of JEV-E protein at different time; (C) Relative expression of MBP protein at different time; (D) Relative expression of NF-H protein at different time.

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Fig 6.

Western blot analysis of Inflammatory cytokines expression levels.

(A) Inflammatory cytokines were determined by Western blot; (B) Relative expression of IL-1β between two groups; (C) Relative expression of IL-6 between two groups; (D) Relative expression of TNF-α between two groups.

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Fig 7.

Brain infection were demonstrated by Immunofluorescence.

A-J: Immunofluorescence analysis of JEV-E protein of mice brains. A through E (Magnification 3x) and F through J(Magnification 30x). A and F: control group. B through E and G through J: The virus group covered different stages.

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Fig 8.

Inflammatory were demonstrated by HE staining of the brains.

A through J:HE staining of mice brains. A through E (Magnification 40x) and F through J(Magnification 80x); A and F: control group. B through E and G through J: The virus group covered different stages. No inflammatory cells infiltrated around the vessels (Yellow arrows in A and B); perivascular cuffing (Yellow arrows in C through E); necrosis lesions and glial nodules (Yellow arrows in I and J).

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