Figure 4.

BMAL1 regulates coagulation factors VII in a direct way. (A) Relative expression of circadian clock genes and coagulation factors at different time points (4 mice/each point). (B) Correlation analysis between BMAL1 and FVII. (C) Plasma concentration of coagulation factors VII and XII at each time point. (D) Motif sequence signature of BMAL1 in the promoter region of coagulation factor FVII. (E) ChIP assays were performed using an antibody against BMAL1, and IgG as negative controls. Specific primers were designed for FVII promoter. (F) Luciferase reporter assays were performed to measure the activities of WT FVII promoter (PGL3- FVII WT), its deletion mutant (PGL3- FVII deletion), its mutant (PGL3- FVII mutation) with mutated BMAL1-binding site and empty vector (PGL3) in Bmal1-overexpression and Bmal1-knockout HepG2 cells. The experiments were performed three times independently. *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001.