Figure 1.

(a) The diagram shows naturally occurring d-flow (lesser curvature, LC) and s-flow regions (greater curvature, GC) in the aortic arch. Also shown is the surgically induced d-flow in the partial carotid ligation model in which three of the four caudal branches of the left common carotid artery (LCA) are ligated, while the contralateral right common carotid artery (RCA) remains untouched in order to serve as an internal control. (b) The scheme shows the intimal RNA extraction step that enables collection of endothelial-enriched RNAs from the mouse carotids. Using an insulin syringe with a 29G needle and 150 μl QIAzol lysis buffer (Qiagen) and carefully flushing the intima with QIAzol lysis buffer into an Eppendorf tube. Intima eluate or the leftover (media+adventitia) are then used for total RNA isolation. (c) Representative image of the ApoE^−/− mouse carotids showing plaque in the LCA after 2 weeks of high-fat diet confirming disease status, while the RCA remains plaque-free. (d) The scheme shows our approach to discover potential, flow-sensitive miRNAs-target gene interactions. Using a simplified hypothesis that the upregulated miRNA will negatively regulate its target gene and vice versa, we initially generated a predicted target gene list for highly differentially regulated miRNAs and then compared this putative target gene list to the list of flow-sensitive mRNAs (that followed an inverse relation with the miRNAs) in the mRNA array. Using this approach, we identified two potential targets for miR-712 as TIMP3 and RECK.