Figure 3.

PAK-independent effects of LPS and EGF. (A) BAECs were treated with LPS or EGF at the indicated concentrations for 30 min and then lysed and analyzed by Western blotting for PAK phosphorylation, Erk phosphorylation, and total Erk as a loading control. Similar results were obtained in 3 independent experiments. (B) BAECs were pretreated the inhibitory peptide described in Figure 7 (PIX-blocking peptide; PBP) or inactive control peptide (Con) at 20 μg/ml. They were then stimulated with LPS or EGF. After 30 min, cells were lysed and analyzed by Western blotting for phosphorylated Erk or total Erk. Similar results were obtained in three independent experiments. (C) BAECs on filters were pretreated with PIX-blocking (PB) peptide or control peptide as in Figure 3B and then stimulated with 10 ng/ml LPS, 10 ng/ml EGF, or 25 ng/ml bFGF as indicated. Permeability across the filter was assayed by movement of HRP as described in Materials and Methods. Values are means ± SD, n = 3.