FIGURE 10.

LYPLA2 inhibition significantly increases endogenous levels of PGE₂-G/PGD₂-G formed in RAW264.7 murine macrophage-like cells. Cells were treated with LPS (1 μg/ml) in the presence or absence of an LYPLA2-specific inhibitor (10 μm) for 6 h in serum-free medium followed by treatment with ionomycin (5 μm) for 45 min. The culture medium was removed, and lipids were extracted for LC-MS/MS analysis for PGE₂-G/PGD₂-G (A) and PGE₂/PGD₂ (B). C, Western blot analysis of untreated and treated RAW264.7 macrophages confirmed the expression of COX-2 and LYPLA2 in cells. β-Actin Western blot verified equal protein loading (10 μg per lane). Data are presented as the mean ± S.D. of triplicate analyses. **, p < 0.01