FIGURE 3.

Murine macrophages generate four 12-KETE-PEs. Murine peritoneal macrophages were isolated from WT mice (8–12 weeks) by lavage with ice-cold PBS. Cells were activated at 37 °C for 15 min with 10 μm A23187, and then lipids were extracted and analyzed using LC/MS/MS as described under “Experimental Procedures.” A, precursor scan for [M − H]⁻ 317.2 shows four major ions generated by murine macrophages. Four prominent ions are circled. B–E, representative chromatograms and MS/MS spectra of lipids detected as parent → 317.2 for the four lipids compared with synthetic standards. Standards were made as described (12) and run under conditions identical to those used for murine macrophage lipid extracts. MS/MS spectra were acquired in ion trap mode at the apex of elution for each lipid. cps, counts/s.