Primary cardiomyocytes were isolated from adult mice (8-12 weeks) following a previous method 24. Briefly, the chest cavity of the anaesthetized mouse was opened to expose the still-beating heart. The heart was perfused with EDTA buffer through left ventricular injection. Afterward, the heart was transferred into a dish and serially perfused with EDTA buffer, perfusion buffer, and collagenase buffer. The tissue was then separated and gently pulled into 1 mm3 pieces using forceps. The cell suspension was passed through a filter, and the cells were collected by gravity with serial reintroduction of calcium buffers to gradually restore calcium concentration. The cells were resuspended in plating media for 1 h at 37 °C in 5% atmosphere. After washing off unattached cells, the remaining cells were incubated in fresh culture media. For hypoxia-reoxygenation (H/R) treatment, cells were subjected to hypoxia (1% O2) for 1 h with or without 10 μM ginsenoside Rb1 (this concentration was chosen based on our previous study 21), followed by reoxygenation. For succinate treatment, cardiomyocytes were incubated in assay buffer (132 mM NaCl, 10 mM HEPES, 4.2 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 25 μM 2-deoxyglucose, 10 mg/L sodium pyruvate; pH 7.4) and subjected to indicated agents in the presence or absence of dimethyl succinate (5 mM) or oligomycin (4 μM) for 2 h.

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