Fig. 3
Transmigration assay across BLECs and HIBCPP cells. a Spontaneous T-cell migration over 2 h across laminin coated 5 μm pore size Millicell® filters is shown. The graph shows the percentage of transmigrated T-cells of healthy donors B (red) and C (blue). Data are displayed as the mean on a superimposed scatter dot plot of 4 independent experiments; 2 experiments for each donor each in triplicates. Statistical analysis: two-way ANOVA followed by Sidak’s multiple comparison (p < 0.05*, p < 0.0001****). b, d, e, g CD4+ T-cell (Th1, Th1*, Th2 and Th17) migration rate across non-stimulated (NS) or 16 h pro-inflammatory cytokine-stimulated (1 ng/mL TNF-α + 20 IU/mL IFN-γ) BLECs and inverted HIBCPP cell monolayers was measured after 8 h transmigration assay. Percentages of transmigrated T cells from peripheral blood of three healthy donors (Donor A–C) and CSF of five MS patients (P-1–P-5) across BLECs (b, e) and inverted HIBCPP cell monolayer (d, g) are displayed. Data are shown as the mean on superimposed scatter dot plot of 5 or 6 independent experiments each performed in triplicates. Statistical analysis: two-way ANOVA followed by Tukey’s multiple comparison test within conditions (between subsets) (p < 0.05*, p < 0.01**, p < 0.001***, p < 0.0001****). Two-way ANOVA followed by Sidak’s multiple comparison test (NS versus stimulated condition) (p < 0.05§, p < 0.01§§, p < 0.0001§§§§). c, f The numbers of CD4+ T-cell (Th1, Th1*, Th2 and Th17) adherent to BLECs monolayer after transmigration assay were displayed. Data are shown as the mean on superimposed scatter dot plot of 3 or 6 independent experiments each performed in triplicates. Statistical analysis: two-way ANOVA followed by Sidak’s multiple comparison test (NS versus stimulated condition) (p < 0.001§§§, p < 0.0001§§§§). Cells used for the representation of the endothelium, the epithelium (HIBCPP cells) and T cells are adapted from Servier Medical Art (http://smart.servier.com/), licensed under a Creative Common Attribution 3.0 Generic License