Figure 6.

Blockade of the PI3K–Akt–GSK3 pathway by specific chemical inhibitors functionally impedes endothelial to monocyte adhesion. HUVEC monolayers were pretreated with LiCl (20 mM), NaCl (20 mM), HGF (100 ng/ml), wortmannin (50 nM), or vehicle before addition of TNF-α (0.1 ng/ml). At different time intervals, Calcein-AM labeled THP-1 cells were applied for monocyte adhesion assay as described in the Materials and Methods. Aliquots of cell lysates were subjected to fluorometric analysis to measure the quantity of adherent monocytes. Values are expressed as mean relative inhibition over HUVEC monolayers pretreated with vehicle±s.d. from three separate experiments. *P<0.05 vs HGF treatment at the same time points.