Fig. 3

Chirality of 15-HETE: ASA triggers 15 R -HETE formation

A549 cells (10⁷ cells/flask) were exposed to IL-1β (1 ng/ml) for 24 hr, treated with vehicle (0.1% v/v ethanol) (□) or ASA (■) for 20 min and then incubated (30 min, 37°C) in HBSS containing arachidonic acid (20 µM) and A₂₃₁₈₇ (5 µM). Products were chromatographed by RP-HPLC (as in Fig. 1B) and the region containing 15-HETE was collected, extracted with chloroform and treated with diazomethane. Chiral analysis was performed with a Bakerbond DNBPG (see Materials and Methods for details). Results are representative of two separate experiments showing similar results. Inset: Ratio between A549-derived 15R- and 15S-HETE in the absence (▨) or presence (■) of ASA.