ChIP-seq for E4BP4 in the liver from transgenic mouse with liver specific E4BP4 overexpression
Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to evaluate the effects of liver-specific E4BP4 overexpression under mouse albumin promoter on the liver glucose and lipid metabolism. Methods: We generated transgenic mice (TG) with liver-specific E4BP4 overexpression, and harvested two livers with one TG mouse each. After cervical dislocation, mouse liver was perfused with a buffer containing Hanks' balanced salt solution, and cross-inked for 30 minutes with 2mM DSG and for 10 minutes with 1% formaldehyde by perfusion, followed by frozen at -80C. Then, Liver was homogenated by gentleMACS Dissociators. Nuclei were isolated and protein-DNA complexes were incubated with antibodies against two kinds of E4BP4 and immunoprecipitated with IgG paramagnetic beads. Results: There were many significant peaks at the promoter/enhancer or intronic region of many genes. Conclusions: E4BP4 possibly regulates the expression of some genes linked to lipid metabolism in the liver. Overall design: ChIP-Seq in liver from transgenic mouse with liver specific E4BP4 overexpression