Lipopolysaccharide-Induced Expression of Microsomal Prostaglandin E Synthase-1 Mediates Late-Phase PGE2 Production in Bone Marrow Derived Macrophages
Figure 6
Selective inhibition of PU.1 didn't affect LPS-induced mPGES-1 expression or late-phase PGE2 production in BMDM.
BMDM was transfected with either PU.1 siRNA or a control siRNA for 36 hrs, and were then treated with or without 1 µg/ml LPS for 16 hrs. The concentration of PGE2 and PGD2 in culture medium was then determined by LS-MC-MC. The protein and mRNA expression in BMDM was determined by Western blot assay or real-time RT-PCR, respectively. A. PU.1 siRNA significantly prevented LPS-induced protein expression of PU.1 in BMDM, but had no inhibitory effect on the protein expression of iNOS, mPGES-1, mPGES-2, or c-PGES (representative blots and the densitometry of iNOS/mPGES-1/PU.1 protein expression were showed from 3 independent experiments). B. Real-time RT-PCR result confirmed that PU.1 siRNA significantly attenuated the PU.1 mRNA expression with or without LPS (16 hrs) treatment in BMDM. C. LPS-induced (16 hrs) PGE2 and PGD2 production in BMDM was not affected by PU.1 siRNA (n = 3).