Fig. 6.

Exogenous ANXA1 enhances endogenous ANXA1 and inhibits activity of the RhoA small GTPase in hCMEC/D3 cells. (A) Treatment of WT hCMEC/D3 cells with 20 μg/mL hrANXA1 for 3 h or 24 h induces an up-regulation in endogenous ANXA1 expression; data are representative of at least three different experiments, performed in triplicate, and are mean ± SEM (*P < 0.05 vs. untreated cells). (B) hCMEC/D3 cells transfected with an antisense sequence to ANXA1 exhibit constitutively increased activity of the small GTPase RhoA, an effect that can be reversed by treatment with 20 μg/mL hrANXA1 for 3 h. Thrombin (Thr; used at 1 IU/mL in serum-free medium for 8 min) was used as a positive control. Data are representative of at least three independent experiments.