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Site-directed mutagenesis was performed using a standard two-step PCR, followed by purification using StarPrep Gel Extraction Kit (D205-04, GenStar, Beijing) and verified by DNA sequencing (Sangon Biotech, Shanghai)

Another circadian clock protein PER2 inhibits the expression of PAI-1 in a CLOCK/BMAL1-dependent manner as an important factor in the development of these metabolic diseases after the circadian clock system is dysregulated (Oishi et al., 2009)