Surgical procedures were carried out according to the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals, and all procedures were approved by the Institutional Animal Care and Use Committee at Temple University Lewis Katz School of Medicine. The IR injury protocol was performed as previously described (34). Briefly, 13- to 15-week-old male mice were anesthetized with 3% isoflurane. The heart was exposed through a left thoracotomy at the level of the fourth intercostal space. A slipknot was inserted around the left anterior descending coronary artery (LAD) at the level of the left auricle with a 6-0 silk suture. After the ligature of the slipknot, the heart was immediately placed back into the intrathoracic space followed by evacuation of pneumothoraces and closure of the muscle and skin. Sham-operated animals were subjected to the same operation, except the suture was passed under the LAD but not tied. After 30 min of ischemia, the slipknot was released, and the myocardium was reperfused for 24 hours unless otherwise denoted.

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