DC adhesion, transmigration, and crawling on lymphatic endothelial monolayers is integrin α4β1/VCAM-1 dependent. In vitro functional assays were performed on inflamed imLEC monolayers. (A–C) Representative FACS plots of imLECs showing ICAM-1 and VCAM-1 expression (red lines) in comparison to isotype control (gray shading). Analysis of adhesion (B) and transmigration (C) of Tln1^−/− DCs as compared with WT DCs. (D) Analysis of the speed of Tln1^−/− and WT DCs. Analysis of adhesion (E) and transmigration (F) of Itgb1^−/− DCs as compared with WT DCs. (G) Analysis of the speed of Itgb1^−/− and WT DCs. Analysis of adhesion (H) and transmigration (I) of WT DCs after treatment with 10 µg/ml anti–ICAM-1, 25 µg/ml anti–VCAM-1, or 10 µg/ml anti–integrin α4 as compared with treatment with the corresponding isotype control. (B, C, E, F, H, and I) Pooled data from three independent experiments are shown. (J) Speed of WT DCs in the presence of 25 µg/ml anti–VCAM-1 or isotype control. (D, G, and J) One representative out of three similar experiments is shown. Each dot represents a single cell. Red line represents the mean. (B–G and J) Unpaired Student’s t test. (H and I) Unpaired one-way ANOVA followed by Tukey post hoc test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001. FSC-A, forward scatter area; Max, maximum; SSC-A, side scatter area.