Effect of aggretin on Akt and ERK1/2 phosphorylation. HUVECs (1 × 106 cells) were cultured in the presence of 0.1 μM aggretin for various times, and then lysed with lysis buffer. Akt or ERK1/2 activation was detected by Western blotting (WB) using antiphosphoAkt pAb or antiphosphoERK mAb coupled with ECL, whereas α-tubulin was used as an internal control. The pattern shown is a representative one of at least 3 similar results (A,C). HUVECs (1 × 106 cells) were cultured in the various concentrations of aggretin for 30 minutes or 24 hours and then lysed with lysis buffer and detected with Western blotting using antiphosphoAkt pAb or antiphosphoERK mAb coupled with ECL, whereas α-tubulin was used as an internal control. The pattern shown is a representative one of at least 3 similar results (B,D).
