Ang-1 diminishes thrombin mediated tyrosine phosphorylation of Occludin.

<p>(<b>A)</b> Serum starved HBMECs were treated with 100nM thrombin for indicated time periods. Cellular proteins were immunoprecipitated with anti-Occludin antibody followed by immunoblotting using anti-PY-20 antibody. Blot was reprobed with anti-Occludin antibody. (<b>B)</b> ECs were stimulated with thrombin (100nM for 15 min) or pretreated with 100ng/ml Ang-1 for 15 min followed by thrombin challenge (100nM for 15 min). Cell lysates were immunoprecipitated with anti-Occludin antibody and probed with PY-20 antibody to detect tyrosine phosphorylation of Occludin. The same blot was reprobed for total Occludin.