Phospho mypt

Human pulmonary artery endothelial cells, small airway epithelial cells (SAEC), and cell culture basal medium with growth supplements were obtained from Lonza (Allendale, NJ). Cells were cultured according to the manufacturer’s protocol and used at passages 5–7. All reagents for immunofluorescence studies were purchased from Molecular Probes (Eugene, OR). PGA2, EP4 agonist CAY10580, and prostaglandin receptor inhibitors were obtained from Cayman (Ann Arbor, MI). The following receptor inhibitors were used: TP inhibitor SQ29548, DP inhibitor BWA868C, IP inhibitor CAY10449, EP4 inhibitors L161982 and GW627368X, FP inhibitor AL8810, and EP1-3 inhibitor AH6809. Rac1 inhibitor NAS23766 was obtained from EMD Millipore (Billerica, MA); PKA peptide inhibitor PKI was from Promega (Madison, WI). VE-cadherin antibody was obtained from Cayman; EP4, Rap1, Rac1, RhoA, ICAM1, and VCAM1 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); β-actin and β-tubulin antibodies were from Sigma-Aldrich (St. Louis, MO); cortactin, phospho-cortactin, VASP, phospho-VASP, phospho-CREB, phospho-MYPT, diphospho-MLC, phospho-NFκB, and IκBα antibodies were obtained from Cell Signaling (Beverly, MA); α-catenin, p120-catenin, and ZO-1 antibodies were from BD Transduction Laboratories (San Diego, CA). Unless specified, other biochemical reagents were obtained from Sigma-Aldrich.