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Real-time quantitative PCR, triglycerides, and Oil Red O staining Intramuscular preadipocytes isolation and plasmid construction Intramuscular preadipocytes of the longissimus dorsi muscle were isolated from calves of Qinchuan cattle at National Beef Cattle Improvement Center Breeding Farm using 0.2% type II collagenase (1148090, Sigma), following the protocol described in our laboratorys previous studies [30,31,32]