Figure 6.

S27D reduces TEM of leukocytes. (A) Primary human neutrophils were allowed to migrate toward 50ng/mL CXCL12 for 1h across TNF-α-pre-treated HUVEC that are cultured on a FN-coated 5-μm pore Transwell filters. Chemotaxis was quantified as described in Methods. Endothelial cells were pre-treated for 30 minutes with DMSO or with S27D, which remained present throughout the experiment. Neutrophil migration was blocked significantly in the presence of the peptide. Data represent mean ± SEM from triplicates. Experiment was repeated three times. *, p<0.05. (B) Differentiated HL60 cells were allowed to migrate across TNFα-treated HUVEC as described under (A). HL60 migration was blocked significantly in the presence of S27D. Data represent mean ± SEM from triplicates. Experiment was repeated three times. *, p<0.01. (C) Differentiated HL60 cells were allowed to migrate across fibronectin-coated Transwell filters toward 50ng/mL CXCL12 for 2 hours in the absence or presence of S27D. Chemotaxis was quantified as described in Methods. Data represent mean ± SEM from triplicates. Experiment was repeated three times. (D) Primary human neutrophils were allowed to migrate as described under A, but as chemokine 10 nM fMLP was used. HUVECs were treated with S27D for 30 minutes, 4.5 hours present during the assay, or washed away after 4.5 hours and left untreated for O/N. S27D inhibited TEM significantly with 4.5 hours but not when washed away. Data represent mean ± SEM. Experiment was repeated three times. *, p<0.05.