Ischemic and non-ischemic hearts from both models were reperfused for a total of 60 min using a modified Krebs-Henseleit buffer containing 1.25 mM Ca2+ and 11 mM glucose. Hearts remained unloaded during the first 10 min. During this time, the left atria of hearts from the in-situ model were cannulated. The perfusion was then switched to working mode for another 50 min. During the 60 min reperfusion, perfusate was recirculating, except the first minute of reperfusion in hearts obtained with the in-situ model, when cardioplegia or blood was washed out and discarded. All hearts were maintained at 37°C with an aortic pressure of 60 mmHg. At the end of the protocol, hearts were immediately frozen in liquid nitrogen and stored at −80°C.

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