FIGURE 8.

Chemical reduction of cell extracts significantly increases detection of 15- or 12-HETE-PE in human monocytes and murine macrophages. A–D, time course of 15-HETE-PE generation by activated human IL-4-treated monocytes with/without SnCl₂ reduction of lipid extracts. Human monocytes were isolated, cultured, and then activated at 37 °C with 10 μm A23187, and at defined time points (15–180 min), some samples were reduced by incubation with 1 mm SnCL₂ for 10 min at 22 °C followed by lipid extraction (n = 3, mean ± S.E.). E–H, time course of 12-HETE-PE generation by activated murine peritoneal macrophages with/without SnCl₂ reduction of lipid extracts. Murine peritoneal macrophages were isolated from WT mice (8–12 weeks) by lavage with ice-cold PBS and pooled. Cells were activated at 37 °C with 10 μm A23187, and at defined time points (15–180 min), some samples were reduced by incubation with 1 mm SnCl₂ for 10 min at 22 °C followed by lipid extraction (n = 3, mean ± S.E.). Error bars represent S.E.