Acetyl coenzyme assay kit

Lactate, pyruvate, and acetyl-CoA levels were separately determined using a Lactate Assay Kit (Jiancheng Bioengineering Institute, Nanjing, China), Pyruvate Assay Kit (Jiancheng Bioengineering Institute), and an Acetyl-Coenzyme Assay Kit (Solarbio, Beijing, China) according to the manufacturer’s protocols. All experiments were performed at least three times.

The levels of glucose, lactate, and pyruvate were separately determined using the Glucose Assay Kit, Lactate Assay Kit, and Pyruvate Assay Kit (Jiancheng Bioengineering Institute, Nanjing, China). The levels of succinate and fumarate were separately measured by using the Succinate Colorimetric Assay Kit and the Fumarate Assay Kit (Sigma, St. Louis, MO). The levels of acetyl-CoA were tested using the Acetyl-Coenzyme assay kit and citrate levels were tested using the Citric Acid Assay Kit (Solarbio, China). The ATP products were measured using the ATP Assay Kit, and the mitochondrial membrane potential was determined using the JC-1 Dye (Beyotime, China). All these measurements follow their respective manufacturer’s protocols and were normalized by cell numbers or the quality of clinical sample. All experiments were performed at least three times.