Fig. 7

CLOCK promotes the expression of ASB9 in GCs. A Sequence analysis of E-box on ASB9 promoter. B Mutated E-box sequences of the luciferase reporter. C and D Luciferase reporter assays of ASB9‐Luc reporter constructs. The wild-type and the mutant pGL3-ASB9 reporters were co-transfected into HEK293T cells with pcDNA3.1(+)/pcDNA3.1(+)-CLOCK plasim, and the fluorescence activity was detected 24 h later. Data are expressed as mean ± SEM (n = 5), ^*P < 0.05, ^****P < 0.0001. E ChIP assay showing recruitment of CLOCK protein to ASB9 E‐box in GCs. The PCR products were analysed on a 2% agarose gel and quantified with densitometry using ImageJ software. IgG and no antibody were used as the negative ChIP control. Data are expressed as mean ± SEM (n = 3), ^*P < 0.05