Thp 1 monocytes

Human aortic endothelial cells were obtained from Cascade Biologics – Invitrogen cell culture (Carlsbad, CA, USA) and were grown in EC culture media MV plus growth supplements from Promo Cell (Heidelberg, DE, USA) supplemented with 100 units/ml penicillin and 100 μg/ml streptomycin. For experiments, 80% confluent ECs were incubated in EC basal media from Promo Cell (2% fetal calf serum, 0.4% EC growth supplements, 90 μg/ml heparin and with 100 U/ml penicillin, and 100 μg/ml streptomycin) for 24 h before treatments. THP-1 monocytes were purchased from the LGC Promochem (Middlesex, UK) and were maintained in RPMI 1640 media supplemented with 10% fetal bovine serum, 1% l-glut, and 1% P/S. The cells were passaged twice weekly in complete media at a ratio of 1:5. All cell lines were maintained in an incubator at 37°C and 5% CO₂. For cell counts and viability determination, the cells were diluted 1:1 with trypan blue and counted using a hemocytometer. 15-Deoxy-Δ^12,14-prostaglandin J₂, biotin-15-deoxy-Δ^12,14-prostaglandin J₂, prostaglandin D₂, and biotin-prostaglandin D₂ were purchased from Cayman Chemical (Ann Arbor, MI, USA). To change the solvent for experiments, prostaglandins were dried down and re-suspended in ethanol at a concentration of 1 mg/ml. For experiments, ethanol was used as vehicle control.

THP-1 monocytes were
purchased from LGC Promochem (Middlesex, U.K.). The Vybrant Phagocytosis
Assay kit (V-6694) was obtained from Molecular Probes. THP-1-Lucia
nuclear factor (NF)-κB cells, heat-killed Listeria monocytogenes
(HKLM), Zeocin, Normocin, QUANTI-Luc, Geneticin (G418), and Hygromycin
were purchased from Invivogen (CA); the Pierce BCA Protein Assay kit
was from Thermo Scientific (Ireland); the Human Pro-Inflammatory multi-Plex
Tissue Culture kit was from MSD (Maryland); and the cytotoxicity detection
kit was from Roche (Germany). Transfecting SMCs with an NF-κB
reporter plasmid pNF-κB-SEAP vector were purchased from Takara/Clontech
(Ireland). HEK-293-transfected and wild-type cell lines were kindly
provided by Actelion Pharmaceuticals Ltd. (Switzerland). For calcium
measurement, sterile plates black-sided with optically clear glass
flat bottoms were purchased from Perkin Elmer (U.K.). LXA₄1 was obtained from Calbiochem (U.K.), and dexamethasone
was from Sigma-Aldrich (Ireland).