We isolated adult mouse CMs according to the previously described procedure 14. MCM and LRP6 Over male mice aged about 8-10 weeks were anesthetized, and the thoracic cavity was opened to expose the heart. The inferior vena and cava-descending aorta were cut and then the heart was immediately flushed by injecting 7-10 mL of EDTA buffer. The heart was immediately digested by sequentially injecting 10 mL of EDTA buffer, 3ml of perfusion buffer, and 50 mL of collagenase buffer into the left ventricle from the aorta. After digestion, the heart tissue was cut into pieces of about 1mm3 and manually dissociated into single cardiomyocyte in 10 % BSA perfusion buffer. The cell suspension was filtered through a 100 μm cell strainer, and the CMs were collected by natural sedimentation for 20 min, and part of the supernatant was centrifuged at low speed to collect non-CMs.

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