*4.4. Multiplex Cytokine Analysis of the Cerebrospinal Fluid*

Multiplex profiling was performed in cerebrospinal fluid (liquor) of experimental and intact animals using the commercially available Bio-Plex Pro™ Rat Cytokine 23-Plex Assay panel, containing the following analytes: G-SCF, GM-CSF, GRO/KC, IFN-g, IL-10, IL-12p70, IL-13, IL-18, IL-1a, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL17A, M-CSF, MCP-1, MIP1a, MIP3a, RANTES, TNFa, and VEGF. Quantitative analysis of target molecules was performed on a Bio-Plex200 System analyzer (BioRad, USA), as recommended by the manufacturer, using Bio-Plex Manager 4.1 software (Bio-RadLaboratories). The statistical processing of obtained results was carried out using linear models implemented in the limma package.

#### *4.5. Statistics*

Statistical data analysis and visualizations were performed using R version 3.5.3 (R Foundation for Statistical Computing, Vienna, Austria). Sample distributions of quantitative values were visualized using box plots, and descriptive statistics are presented as: (Median [1st quartile; 3rd quartile]). The Kruskal–Wallis test was used to compare morphometric and immunofluorescent staining data between experimental groups when a null hypothesis was rejected; we used Dunn's-test as the post hoc method. Differences were considered statistically significant where *p* < 0.05.
