Figure 5. Inhibition of ADAM10 decreases transendothelial migration of T cells.

A) HUVECs were grown to confluence on 3 μm pore size transwell filters and pretreated with DMSO, broad-spectrum metalloproteinase inhibitor GM6001 (10 μM), GI254023X (10 μM) or GW280264X (10 μM) for 1 hour. After medium changes in the upper chamber, HUVECs were incubated with 5×10⁵ T cells (PHA blasts) per filter. T cell transmigration was measured by counting cells in the lower chamber after 24 hours. Data are expressed as mean±SEM; n=3 independent experiments. ^*P<0.05 vs DMSO treated cells. B) Either T cells or HUVECs were transfected with control siRNA (1, insert) or ADAM10 siRNA (2, insert) and analyzed for T cell transmigration. The downregulation of ADAM10 (A10, insert) expression was confirmed using immunoblot analyses. p: precursor of ADAM10; m: mature form of ADAM10. Data are mean±SEM; n=3 independent experiments. ^*P<0.05 vs mock transfected cells.