LPS induces increased expression of iNOS/COX-2/mPGES-1/PU.1 in BMDM.

<p>Primary cultured mouse BMDM was treated with or without 1 µg/ml LPS for various time points from 2 to 24 hrs. <b><i>A</i></b>. The protein expression of targeted proteins in BMDM cell lysates was detected by Western blot assays. Representative blots and the densitometry of iNOS/COX-2/mPGES-1/PU.1 protein expression are showed from 5 independent experiments. Protein expression of COX-2 and iNOS was detected within 2 to 4 hrs post-LPS treatment; whereas increased protein expression of mPGES-1 and PU.1 was detected after 8 hrs of LPS treatment. In contrast, the protein expression of COX-1, mPGES-2, c-PGES or H-PGDS was not affected by LPS treatment. The protein loading in each experiment was normalized by β-actin. <b><i>B–G</i></b>. The LPS-stimulated (16 hrs) mRNA expression of COX-2 (<b><i>B</i></b>), iNOS (<b><i>C</i></b>), PU.1 (<b><i>D</i></b>), mPGES-1 (<b><i>E</i></b>), mPGES-2 (<b><i>F</i></b>), or c-PGES (<b><i>G</i></b>) in BMDM was confirmed by real-time quantitative RT-PCR. The mRNA of each gene was normalized to the β-actin mRNA expression level in the same sample. The results shown are mean of at least 4 independent experiments for each gene, * <i>p</i><0.05 vs. control.