Updated project metadata. TNFα is a cytokine with multiple biological effects. It activates cascading signal pathways by binding to receptors on the cell membrane, regulating processes such as cell growth, differentiation, apoptosis, and inflammatory responses. The formation of skeletal muscle primarily relies on processes such as myoblasts proliferation, migration, differentiation, cytoskeletal rearrangement, and myotube fusion, which constitute a complex and coordinated process regulated by multiple genes. While research on TNFα has primarily focused on immunology, some studies suggest that TNFα also plays a physiological role in myogenesis, muscle repair, diseases, and atrophy.We isolated the nuclei (N) and cytoplasm (C) of flox and TNFα-CKO primary myoblasts in the early stages of differentiation and attempted to identify new targets in the TNFα signaling pathway.