Reversal of angiostatin inhibition of AKT phosphorylation with excess HGF.

HUVECs in serum-free medium were untreated (lane 1) or treated with angiostatin (3 μM) and increasing concentrations of HGF: 0.5 ng/mL (lane 2), 10 ng/mL (lane 3), 25 ng/mL (lane 4), 50 ng/mL (lane 5), and 100 ng/mL (lane 6). After a 5-minute incubation, cell lysates were prepared and immunoblotting for phospho-AKT was performed as described. The blot was stripped and re-probed for total Akt.