FIGURE 4.

Knockdown of TRAP1 alleviates EAE pathogenesis in mice. Mice were injected to recombinant lentiviruses carrying the promoter of GFAP and TRAP‐shRNA or ctrl‐shRNA sequence, for 7 days, prior to MOG_35–55 immunization for 21 days (n = 10 mice per group). (a) The clinical scores of EAE mice by TRAP‐shRNA and ctrl‐shRNA treatment. (b) Western blot assay was employed to detect the protein levels of TRAP1, Smad4, p‐Smad2/3, Smad2/3, and GFAP in the spinal cords. (c and d) The expression level of TNF‐α, CXCL10, and MCP‐1 in the spinal cords and peripheral blood of the TRAP‐shRNA and ctrl‐shRNA mice were measured by real‐time PCR assay and ELISA, respectively. The data are represented as mean ± SEM. ***p < .001 versus NC group; ^# p < .05 and ^## p < .01 versus ctrl‐shRNA group (n = 10 mice/group). (e) Hematoxylin and eosin (H&E) staining was employed to evaluate the infiltrations of inflammatory cells in spinal cords (scale bars, 50 μm). (f) Luxol fast blue (LFB) staining was used to investigate the medullary sheath damages from spinal cords (scale bars, 50 μm). Red box areas in the upper rows are presented enlarged underneath. (g) Electron microscope (EM) was employed to observe the severe disruption or mild loosening of the medullary sheath in spinal cords. Red arrow indicates the changes of the medullary sheath. Scale bars, 1 μm