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Figure 1 Schematic representation of repetitive ischemia and reperfusion (I/R) protocol and adeno-associated virus (AAV)-2-mediated intramyocardial gene transfer. (A) Illustration of the experimental protocol for repetitive ischemia. AAV-HO-1 was administered five weeks in advance of the injury, followed by left anterior descending coronary artery (LAD) occlusion for 15 min and release performed daily for five days. Twelve days after the last ischemic episode, animals were subjected to echocardiography followed by tissue harvest and analyses. (B) Schematic diagram of the sites of injection (indicated by filled circles). (C) Representative electrocardiogram (ECG) at basal showing normal tracing and showing an increase in ST-segment following LAD ligation. (D) Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) detection of HO-1 transcript in left ventricle five weeks after gene transfer. (E) Western blot analysis of distribution of HO-1 protein in injected (ischemic) and non-injected (remote) areas of the left ventricle.