Figure 6.

Eicosanoid synthesis, eicosanoid biosynthetic enzyme expression, and cAMP levels in AMs from WT (open bars) and l/l (solid bars) mice. AMs from WT (open bars) and l/l (solid bars) mice were stimulated with heat killed K. pneumoniae (K.p.) opsonized with specific immune serum for 1 h and cysteinyl-leukotrienes (cysLT) and prostaglandin E₂ (PGE₂) were determined by ELISA as described in the Materials and Methods section (A). Immunoblots for 5-lipoxygenase (5-LO), cyclooxygenase-1 (COX-1), microsomal prostaglandin E synthase-1 (mPGEs-1), and glyceraldehydes-phosphate-dehydrogenase (GAPDH) (protein loading control) in AM lysates from a single experiment that is representative of a total of 3 independent experiments. Bars represent the mean arbitrary densitometric units ± SEM of immunoblots from 3 separate experiments (B). AMs cultured with and without indomethacin (Indo) (10 µM) for 30 min were stimulated with or without heat killed K. pneumoniae (K.p.) opsonized with immune serum for 1 h and the cell culture media was assayed for PGE₂ (C) and AM lysates were assayed for cAMP as described in the Materials and Methods section (D). * P<0.05 vs WT using the students t-test.