Mitochondrial Membrane Potential Assay

To detect mitochondrial membrane potential, cardiomyocytes were incubated with JC-1 (Beyotime, Shanghai, China) for 20 min at 37 °C. Cardiomyocytes were analyzed within 1 h using flow cytometry (CytoFLEX LX, Beckman, USA). All data were analyzed using EXPO32 ADC analysis software (Beckman, Washington, USA). For flow cytometry analysis, unstained cells were used as controls to identify the forward scatter/side scatter (FSC/SSC) gates used in the same experiments. A total of 10,000 cells was detected in each sample [30 (link)].

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Jiang M.Y., Man W.R., Zhang X.B., Zhang X.H., Duan Y., Lin J., Zhang Y., Cao Y., Wu D.X., Shu X.F., Xin L., Wang H., Zhang X., Li C.Y., Gu X.M., Zhang X, & Sun D.D. (2023). Adipsin inhibits Irak2 mitochondrial translocation and improves fatty acid β-oxidation to alleviate diabetic cardiomyopathy. Military Medical Research, 10, 63.

Publication 2023

CytoFLEX LX EXPO32 ADC analysis software

Corresponding Organization : Chinese PLA General Hospital

Other organizations : Xijing Hospital, Air Force Medical University

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Variable analysis

independent variables

Incubation with JC-1

dependent variables

Mitochondrial membrane potential

control variables

Cardiomyocytes incubation temperature of 37 °C
Duration of JC-1 incubation (20 min)
Analysis within 1 h using flow cytometry
Unstained cells used as controls to identify the forward scatter/side scatter (FSC/SSC) gates

controls

Unstained cells used as negative control

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