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. Author manuscript; available in PMC: 2010 Jul 1.
Published in final edited form as: Free Radic Biol Med. 2009 Apr 17;47(1):62–71. doi: 10.1016/j.freeradbiomed.2009.04.016

Fig. 3.

Fig. 3

Metabolism of AA by freshly isolated and cultured neutrophils. Freshly isolated neutrophils (A,C) or neutrophils that had been cultured for 24 h in RPMI/10% FBS (B,D) were suspended in PBS and incubated for 2 h (A,B) or various additional times (C,D) with AA (20 μM) and A23187 (5 μM). The products were measured by RP-HPLC as described in Materials and Methods (A,B) using a gradient prepared from solvents A (water), B (acetonitrile) and C (methanol), all containing 0.02% acetic acid, as follows: 0 min: 60%A, 30%B, 10%C; 42 min: 10%A, 38%B, 52%C. The flow rate was 1 ml/min. PGB2 (100 ng) was used as an internal standard. The upper and lower tracings show absorbance at 235 and 280 nm, respectively. The time courses for the formation of 5-HETE (5h, ○), 5-oxo-ETE (5o, •), LTB4 (B4, ▲), and 20-hydroxy-LTB4 (hB4, Δ) by freshly isolated neutrophils (C) and neutrophils cultured for 24 h (D) are also shown (n = 5). Other abbreviation in panels A and B are: dh, 5,20-diHETE; oh, 5-oxo-15-HETE; 15h, 15-HETE.