Fig. 5
Effect of Ki16425 on BBB disruption in spinal cords of EAElow mice. A–K Lumbar spinal cords (n = 3 per group) were dissected from the sham, EAElow, EAElow + Ki16425 (15 and 30 mg/kg), and Ki16425 (30 mg/kg) groups at day 19–20 after immunization. A, F, and I Cryosections (n = 3 per spinal cord) were subjected to immunofluorescence staining with anti-albumin (upper panel in A), anti‐IgG (lower panel in A), anti-PECAM-1 (upper panel in F), anti‐GFAP (lower panel in F), anti-occludin (upper panel in I), or anti-claudin-5 antibody (lower panel in I). Their images were captured by confocal microscope and the levels of relative intensity were measured (B and C). D and E Lysates from lumbar spinal cords were analyzed by Western blot to investigate protein expression of PECAM-1 (D) and GFAP (E). G, H, J, and K The lysates (n = 3 per group) were analyzed by real-time PCR to measure the degrees of occludin (G) and claudin-5 (H) to represent tight junctions and ICAM-1 (J) and VCAM-1 (K) to represent cell adhesion molecules. Bar = 100 µm. Quantified data are expressed as mean protein level or fold induction ± SEM. (ANOVA test; #p < 0.05 versus sham group; *p < 0.05 and **p < 0.01 versus EAElow group