Gsk0660

Human umbilical vein ECs (HUVECs) were isolated as described previously [10 (link), 26 (link)]. HUVECs from 3 to 4 cords were pooled for experiments, and passage 2–3 HUVECs were used for all experiments. HUVEC culture medium included M199 media (Invitrogen, Carlsbad, CA) supplemented with 10 % fetal bovine serum (FBS) (Invitrogen), 30 µg/ml endothelial cell growth factor supplement (ECGF) (BD Biosciences, San Jose, CA), 100 µg/ml heparin sodium (Sigma, St Louis, MO), 1 % sodium bicarbonate (Invitrogen), 1 % Glutamax™-1 (Invitrogen), 1 % penicillin/streptomycin (Invitrogen), 0.25 % fungizone (Invitrogen), and 0.25 % gentamicin (Invitrogen). Cells were cultured at 37 °C with 5 % CO₂ and humidity. In some experiments, cells were treated with VEGF-A 165 (R&D Systems, Minneapolis, MN) or chemically defined lipid mixture 1 (Sigma). For induction of apoptosis, HUVECs were cultured in medium containing 5 % FBS and no ECGF, or treated with TNFα (40 ng/ml) for 24 h. PPARδ agonist GW0742 and PPARδ inhibitor GSK0660 were purchased from Cayman Chemicals (Ann Arbor, MI) and R&D Systems, respectively.