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Image Search Results
Journal: Arteriosclerosis, Thrombosis, and Vascular Biology
Article Title: PAR4 Inhibition Reduces Coronary Artery Atherosclerosis and Myocardial Fibrosis in SR-B1/LDLR Double Knockout Mice
doi: 10.1161/ATVBAHA.123.319767
Figure Lengend Snippet: RAG8 treatment impairs PAR (protease-activated receptor) 4–mediated platelet activation. A , SR-B1 (scavenger receptor class B type I)/LDLR (low-density lipoprotein receptor) double knockout (DKO) mice fed the high-fat, high-cholesterol diet containing cholate (HFCC) for 20 days and treated with the RAG8 or the inactive control SRQ8 pepducins were anesthetized, and their tails were transected across the vein ≈0.5 cm from the tip and placed in a cuvette with warm saline. The time to cessation of bleeding was monitored. Each data point represents an individual mouse (n=12, 14). B and C , SR-B1/LDLR DKO mice were fed the HFCC for 14 days and treated with the RAG8 or the inactive control SRQ8 pepducins (n=4 each). Platelets were prepared from blood collected retro-orbitally from anesthetized mice. Platelet activation was measured by treatment of platelets with either thrombin or a synthetic PAR4-AP (PAR4 agonist peptide) by flow cytometry after immunostaining with either ( B ) a FITC (fluorescein isothiocyanate)-labeled Wug-E9 antibody (for P-selectin) or ( C ) a PE (phycoerythrin)-labeled JON/A antibody for active GP IIb/IIIa (glycoprotein). Resting platelet activation levels do not differ between control SRQ8- and RAG8-treated SR-B1/LDLR DKO mice. Units are expressed as median fluorescence intensity (MFI). Bars represent means, and error bars represent SEMs. Data in A passed the normality test and were analyzed by Student t test. Data in B and C were analyzed by 2-way ANOVA with Sidak multiple comparisons post hoc test. **** P <0.0001.
Article Snippet: For pepducin-treated mouse groups, platelets (1×10 6 washed mouse platelets per reaction) were resting or activated with 1 U/mL α-thrombin (HT 1002A; Enzyme Research Laboratories, South Bend, IN) or 300 μM
Techniques: Activation Assay, Double Knockout, Control, Saline, Flow Cytometry, Immunostaining, Labeling, Fluorescence