Fig. 1From: Aspirin-Triggered Lipoxins (15-epi-LX) Are Generated by the Human Lung Adenocarcinoma Cell Line (A549)–Neutrophil Interactions and Are Potent Inhibitors of Cell Proliferation PGHS and LO expression and monohydroxy products analysis in A549 cells (A) PGHS and LO expression in A549 cells. Cells were grown for 24 hr at 37°C in T-75 cm2 flasks in the presence or absence of IL-1β (1 ng/ml). Extracted total RNA (1 µg) was taken for RT and PCR using specific oligonucleotides for PGHS-1 and -2, 15-, 12-, and 5-LO, and GAPDH. Radioactive bands were quantified directly by phosphorimager analysis, normalized to the expression of GAPDH, and expressed as fold increase in mRNA levels after exposure to IL-1β. Inset: 15-LO mRNA expression in human lung tissue and peripheral blood monocytes (PBM). ND, not detected. (B) RP-HPLC profile of [3H]-labeled mono-HETEs from permeabilized IL-1β-treated A549 cells (1.5 µ 106 cells/ml) and exposed to [3H]-arachidonic acid (0.25 µCi/ml) plus unlabeled arachidonic acid (20 µM) for 20 min at 37°C. Products were extracted and chromatographed using a linear gradient of methanol:H2O:acetic acid (65:35:0.01; v/v/v) and methanol:acetic acid (99.99:0.01, v/v) at a flow rate of 1.0 ml/min. Arrows denote co-chromatography of synthetic standards.Back to article page