FIGURE 3.

(A) Subcellular localization of 5-LOX by immunostaining. PMNLs (2 × 10⁶/mL HBSS/HEPES) were incubated without additived (vehicle), with nonopsonized S. typhimurium (25:1 bacteria per cell ratio), with 0.1 µM fMLP or under sequential stimulation with these agents at intervals of 20 min. The final incubation with fMLP in this case lasted 5 min. At the end of the incubation, the cells were fixed and stained for 5-LOX (green). Samples were also stained for DNA with Hoechst (blue). (B–E). Scanning electron microscopy appearance of neutrophils in co-incubation with bacteria and fMLP. PMNLs (10⁶/ml) in HBSS/HEPES medium were plated on coverslips and incubated for 20 min at 37°C in the medium (B, C), or with nonopsonized S. typhimurium (25:1 bacteria per cell ratio) (D, E), followed by adding fMLP (0.1 µM) for 5 min (C, E).