Characterizing PAFR Signaling in Human Cells

All chemicals were obtained from Sigma-Aldrich (St. Louis, MO) unless indicated otherwise. The culture media was purchased from Mediatech, Inc. Manassas, VA). The PAFR agonist, carbamyl-PAF (CPAF), and PAFR antagonist (WEB2086) were from Cayman Chemicals Co. (Ann Arbor, MI). The human IL-8 ELISA kit was purchased from R&D Systems, Inc. Minneapolis, MN. The PAFR-expressing (KBP) cells were generated from PAFR-negative KB cells via transduction with the MSCV2.1 retrovirus encoding the human leucocyte PAFR, and PAFR-deficient (KBM) cell by transducing with the MSCV2.1 retrovirus vector alone. These human cell lines were cultured in DMEM media supplemented with 10% FBS, 6mM L-glutamine, and a 100μg/ml mixture of penicillin and streptomycin as per our previous reports. ^{29 (link)–30 (link)}