Cardiomyocytes were washed twice with cold PBS and lysed with lysis buffer containing phosphatase inhibitors for 30 min at 4°C. Samples were centrifuged at 13,000 rpm for 15 min at 4°C, and the supernatant was collected for use in the firefly luciferase-based ATP assay according to the manufacturer’s instructions. The luminescence signal was measured using a luminometer (Tecan Infinite M200 Pro) at 560-nm absorbance. The firefly luciferase-based ATP determination kit (Molecular Probes) was used to measure mitochondrial ATP content. Each assay was performed at least three times (Yan et al., 2019 (link)).
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