Fig. 5
EGFL7 treatment decreases expression of MCAM by BBB-ECs. a–c Expression of cell adhesion molecules ICAM-1, MCAM, and VCAM-1 was assessed by immunofluorescence on frozen sections (cerebellum, brainstem, and spinal cord) from representative EAE mice treated with rmEGFL7 (lower panels, spinal cord) or vehicle (upper panels, spinal cord) as described. Left column: ICAM-1 or VCAM-1 (red), DAPI (blue). Middle column: MCAM (green), DAPI (blue). Right column: merge (yellow). Scale bar = 50 μm. n ≥ 5 sections/mouse, from n = 5 mice/group. b, c Pixel intensity and relative pixel intensity of ICAM-1, MCAM and VCAM-1 as calculated using ImageJ software. d Number of cells in bottom chamber after 16 h migration over inflamed human BBB-ECs (modified Boyden chamber assay), following 24 h exposure to rhEGFL7 10 μg/ml or vehicle. One million activated CD4CD45RO T lymphocytes were seeded. n = 5 donors, two different preps. Statistical analysis performed by Mann-Whitney U-test (b, c) and by Wilcoxon matched-pairs signed rank test (d). *p < 0.05. All data presented as mean ± SEM
