Files in this Data Supplement:

• Figure I - (Microsoft PowerPoint file) (600 KB). Expression of mPGES-1 in aorta of Ang-II treated LDLR^-/- (A, B) and mPGES-1^-/- LDLR^-/- (C, D: negative controls) mice. OCT frozen sections were stained for mPGES-1. Representative sections from two different mice of each phenotype are displayed.
• Figure II - (Microsoft PowerPoint file) (670 KB). Expression of COX-1 and COX-2 in aorta of LDLR^-/- and mPGES-1^-/- LDLR^-/- mice. OCT frozen sections of aorta from untreated and Ang-II treated LDLR^-/- and mPGES-1^-/- LDLR^-/- mice were stained for COX-1 or COX-2 as indicated.
• Figure III - (Microsoft PowerPoint file) (210 KB). Aortic expression of angiotensin receptors. No statistical significance was detected by RT-PCR for Ang II receptors, AT1a, AT1b and AT2, between the Ang II treated LDLR^-/- and mPGES-1^-/- LDLR^-/- mice (A). Western blot showed no difference in AT1 and AT2 between the two AngII treated groups (B). 20 μg aortic protein was loaded each lane. Rabbit anti-mouse AT1 and AT2 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) were used as primary antibody with dilution of 100 and 250 folds, respectively. HRP labeled anti rabbit secondary antibody diluted by 2000 folds was used. Rabbit anti-mouse beta-actin antibody (Santa Cruz Biotechnology, Santa Cruz, CA) was diluted 5000 fold.
• Figure IV - (Microsoft PowerPoint file) (661 KB). MMP activity assay by gelatin zymography. MMP2 activity was depressed by deletion of mPGES-1. Cleaned and frozen aortae from 6 LDLR^-/-s and 5 mPGES-1^-/- LDLR^-/-s were homogenized in 150 μl lysis buffer (1% Triton X 100, 0.1%SDS in PBS) with TissueLyser(Qiagen, Valencia, CA), two rounds at 30 Hz for 3 min. Aortic homogenate from each aorta was centrifuged at 10K rcf for 5 min and supernatant containing 20 μg protein, as quantified by Biorad Protein Assay, was loaded onto 10% Zymogram (Gelatin) Gel (Invitrogen, Carlsbad, CA). Protein molecular weight marker was loaded in lane M. Following electrophoresis, the gel was washed briefly with water, re-natured in Novex Zymogram Renatureing Buffer (Invitrogen, Carlsbad, CA) for 30 min, and then incubated overnight with Novex Zymogram Developing Buffer (Invitrogen, Carlsbad, CA) at 37°C. The gel was washed three times with water for 5 min. The zones of lysis were visualized by staining with 0.5% Coomassie blue R-250, followed by a 1 hour wash with water. Quantitative results, shown in the bar graph is calculated from the gel image by Image J 1.38x software. Integrated density was used for each band and normalized to the protein loaded. *: p< 0.05