Increased transmigration of MLN lymphocytes across endothelial monolayers expressing high levels of JAM-2.
(A) Nontransfected tEnd.1 cells and cells transfected with either JAM-1 or JAM-2 were cultured in Transwell culture inserts in 24-well plates. Forty-eight hours later, the cultures were washed and 1 × 106 splenic or MLN lymphocytes were added to each filter and incubated for 3 hours. The chemokine mouse SDF-1 was added to the lower chamber. At the end of the assay, transmigrated cells were collected from the lower chamber and counted by light microscopy. The results are expressed as the percentage of cells transmigrated calculated from the mean of triplicate filters (± SD) and are representative of at least 3 independent experiments. (B,C) A time course of transmigration was carried out on monolayers of tEnd.1 cells, either nontransfected (open bars) or transfected with JAM-2 (hatched bars) as for panel A, except that transmigrated lymphocytes were collected at the times indicated. Two representative experiments are shown *P < .05. **P < .01.
