Figure 7.

Time- and concentration-dependent biosynthesis of 15-LO metabolites from 1-LG, LEA, and LA by human neutrophils. Pre-warmed human neutrophils suspensions (37 °C, 5 × 10⁶ cells/mL in HBSS containing 1.6 mM CaCl₂) were treated with 3 μM of (A) 1-LG, (B) LEA, or (C) LA for the indicated times. (D–F) Neutrophils were treated with increasing concentration of (D) 1-LG, (E) LEA, or (F) LA for 5 min. In experiments involving 1-LG, cells were pretreated with 1 µM MAFP for 5 min prior to the addition of 1-LG. Incubations were stopped by the addition of one volume of cold (−20 °C) MeOH containing the internal standards. Samples then were processed for LC-MS/MS analyses as described in methods. Data are the mean (±SEM) of 4 independent experiments.