Figure 3

Slit2 mediates chemokinetic and chemotactic behaviour on endothelial cells while serum exclusively mediates chemotaxis. A shows the migration of control lacZ siRNA and robo4 siRNA transfected endothelial cells to AP and AP-Slit2N (25 ng/ml) fusion proteins in a Boyden chamber assay. The data here is consolidated from three independent experiments with each experiment performed with samples in triplicate. B shows the AP activity in lysates prepared from untransfected (UT), AP and AP-Slit2N treated control and robo4 siRNA transfected endothelial cells. C shows migration assay for control lacZ and robo4 siRNA transfected cells to Serum or AP-Slit2N in either upper (U), lower (L) or both chambers as indicated. Error bars in A (n = 3), and B (n = 3) represent SD while in C represent SEM (n = 4). D shows pulldown analysis of Cdc42-GTP levels in AP and AP-Slit2N (25 ng/ml) treated endothelial cell lysates for 5 and 15 minute respectively. + indicate addition of the reagent on the left, pd: pulldown, total: total Cdc42 protein in lysates.