The arterial blood was collected from mice left ventricle and human radial artery respectively, the erythrocytes were subsequently lysed by lysing buffer (BD Biosciences, USA). Specifically, 10 times the volume of the lysing buffer was added into the arterial blood. Gently vortex each tube immediately and incubate at room temperature, protected from light, for 15 min. Then the tubes were centrifuged 3000X g for 10 min. The supernatant was carefully aspirated and repeat the above steps. Then mitochondria were isolated with a cellular mitochondria isolation kit (Beyotime Biotechnology, China) according to the manufacturer’s protocol.
Briefly, the pellet resuspended in 0.5 mL mitochondria isolation buffer and grinded with three-dimensional freezing grinder under the frequency of 60 Hz. Then the tubes were centrifuged 600X g for 10 min. The mitochondrial solution was collected carefully without disturbing pellet. Finally, the mitochondria were obtained after centrifugation 600X g for 10 min. The isolated mitochondria were then counted by a Vi-cell XR system (Beckman coulter, Inc. USA).
Briefly, the pellet resuspended in 0.5 mL mitochondria isolation buffer and grinded with three-dimensional freezing grinder under the frequency of 60 Hz. Then the tubes were centrifuged 600X g for 10 min. The mitochondrial solution was collected carefully without disturbing pellet. Finally, the mitochondria were obtained after centrifugation 600X g for 10 min. The isolated mitochondria were then counted by a Vi-cell XR system (Beckman coulter, Inc. USA).