sE-selectin induces HMVEC chemotaxis through Src and PI3K pathways.
(A) HMVECs (1 × 106 cells/mL) in the lower chamber were incubated with various concentrations of sE-selectin in the upper chamber for 2 hours at 37°C. PBS was used as a negative control and bFGF (60 nM) as a positive control. Results are expressed as the number of cells migrating through the membrane per well ± SEM from 3 independent experiments. sE-selectin showed a dose-dependent increase in HMVEC chemotaxis compared with negative control PBS (P < .05). *Represents a significant difference (P < .05) between the sE-selectin and negative control PBS. (B) For inhibition studies, HMVECs were pretreated with the respective inhibitor or inhibitor combination (10 μM PP2, LY294002, or PD98059; or 50 ng/mL PT) for 2 hours at 37°C and then assayed in 48-well chemotaxis chambers in response to 50 nM of sE-selectin. Results are expressed as the number of cells migrating through the membrane per well ± SEM from 6 independent experiments. PT in the figure represent pertusis toxin. *Represents a significant difference (P < .05) between the respective groups. sE-selectin induced HMVEC chemotaxis predominantly through the Src and PI3K pathways.
