FIGURE 5.

Plasmin-induced PKC activation requires its protease activity. A, CMECs were treated with either active or d-Val-Phe-Lys chloromethyl ketone (VPLCK)-inactivated plasmin for the indicated time periods. Phospho-PKC, -ERK1/2, and -p38 were evaluated by immunoblot using specific antibodies. β-Tubulin served as a loading control. B, CMECs were treated with plasmin at 0.3 unit/ml for 7.5, 15, or 30 min. A2, p11, and CD47 levels were examined by surface biotinylation followed by immunoblot analysis using rabbit polyclonal anti-A2 directed at the N-terminal tail domain of A2, monoclonal anti-p11, and goat anti-CD47 IgGs. Total A2, p11, and phospho-PKC were assessed by immunoblot analysis.