Fig. 6

cg09897064 methylation impairs the CEBPA binding to promoter domain of ZBP1. A. CD14⁺ mononuclear macrophages, transfected with both the pGL3 luciferase vector and the pGL3 control vector, were washed and subjected to a luciferase activity assay after 24 h.The relative luciferase activity was then measured in triplicate wells, with both unmethylated and methylated promoter regions present. B. Motif analysis of ATACseq data revealed that CEBPA binds to the promoter domain of ZBP1, as indicated by the identified binding sites. C. An electrophoretic mobility shift assay(EMSA)was employed to evaluate the CEBPA's adhesion to both unmethylated and methylated promoter areas. D. The diagram illustrates how cg09897064 methylation regulates ZBP1 transcription. It shows that cg09897064 methylation impairs the binding of CEBPA (Transcription factor, TF) to the promoter domain of ZBP1. Statistical significance was established through appropriate tests(*P < 0.05,**P < 0.01,***P < 0.001)for the mean ± standard deviation of the presented data