A and B- Pulmonary EC were stimulated with OxPAPC (10 μg/ml), thrombin (0.1 U/ml, 10 min), or vehicle for various time periods followed by immunoprecipitation of: A - p120-catenin and B - or ZO-1. Afadin, VE-cadherin, p-120 catenin, and ZO-1 content in the immunoprecipitates was detected using specific antibodies. Equal protein loading was confirmed by membrane reprobing with p120-catenin (A) or ZO-1 (B) antibodies. Results are representative of four independent experiments. C - HPAEC transfected with Rap1-specific siRNA or non-specific RNA were treated with OxPAPC (10 μg/ml) or vehicle for 30 min followed by immunoprecipitation of p-120 catenin or ZO-1. Afadin content in the immunoprecipitates was determined using specific antibodies. Results are representative of three independent experiments.