For detecting the concentrations of triglyceride, ATP, and ROS, bovine intramuscular preadipocytes were seeded in 24-well plates the day before transfection. pCDNA-ACOX1, pCDNA-3.1(+), Si-ACOX1 and NC were transfected into confluent (~80%) cells, respectively. After 24–48 h, the concentrations of triglyceride and ATP in the lysates of cells were measured with commercial kits (Applygen (Beijing, China) and Beyotime (Shanghai, China), respectively) following the manufacturer’s instructions, and normalized to the protein content (µ mol/mg protein) using the BCA assay kit (Thermo Scientific). ROS were measured using the reactive oxygen species assay kit (Beyotime) following the manufacturer’s protocol.

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