Fig. 5

4-EG mitigates BBB disruption, spinal cord pathology, and MMP3/9 expression in EAE. a At day 12–13 post-immunization, vehicle- and 4-EG (100 mg/kg)-treated C57BL/6 EAE mice were subjected to i.v. injection of Evans blue. Two hours after Evans blue injection, EAE mice were sacrificed, and the brains and spinal cords were harvested to analyze Evans blue leakage. Representative brain and spinal cord images of three vehicle- and 4-EG-treated EAE are shown, and the amount of Evans blue leakage in the brain and spinal cord tissues was also quantified (n=7/group). b The lumbar regions of the spinal cord harvested from vehicle- and 4-EG-treated C57BL/6 EAE mice were subjected to H&E staining to determine peripheral immune cell infiltration. Representative H&E images of the coronal spinal cord sections are shown. Scale bars, 200 μm (×10, left image), 50 μm (×20, right image). The areas of cell infiltration were quantified (n=6/group). c The brain and spinal cord tissues were harvested from vehicle- and 4-EG-treated C57BL/6 EAE mice (n=5/group) and then subjected to western blot analysis to measure MMP3 and MMP9 expression. Data are representative of two independent experiments (n=5/group per experiment). d The lumbar regions of the spinal cord from vehicle- and 4-EG-treated C57BL/6 EAE mice were subjected to IHC to determine MMP9 and Iba1 expression. Representative images of MMP9 and Iba1 expression are shown. The fluorescence intensity of MMP9 was also quantified (n=7/group). Scale bar, 50 μm (×20). Statistical significance was determined as *p<0.05, **p<0.01, and ***p<0.001 by unpaired t test