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Besides genetic disorders, skeletal muscle atrophy mainly occurs as a consequence of underlying conditions such as prolonged inactivity, aging, and metabolic diseases, ultimately contributing to the risk of disability. Disturbances in cellular and molecular mechanisms involved in proteolysis and protein synthesis underpin muscle fiber shrinkage and decreased muscle fiber diameter. Stress-induced primary myotube culture is an established model for studying muscle atrophy. An in vitro model is an essential criterion in establishing preliminary data in a cell-autonomous manner that can later be validated using in vivo models. Here, we describe protocols for the isolation, culture, and differentiation of primary murine myotubes and the induction of myotube atrophy using dexamethasone, a synthetic corticosteroid. We further elaborate the procedure to validate degenerative parameters, such as assessing muscle fiber diameter, expression of muscle atrophy genes, and protein synthesis status under dexamethasone treatment. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Isolation and culture of primary myoblasts from rat or mouse pups Support Protocol 1: Preparation of coated tissue culture ware Support Protocol 2: Subculture of myoblasts Basic Protocol 2: Induction and assessment of myotube atrophy.

Sirtuin 6 (SIRT6) is a member of the mammalian sirtuin family with deacetylase, deacylase, and mono-ADP-ribosyl-transferase activities. It is a multitasking chromatin-associated protein regulating different cellular and physiological functions in cells. Specifically, SIRT6 dysfunction is implicated in several aging-related human diseases, including cancer. Studies indicate that SIRT6 has a tumor-specific role, and it is considered a tumor suppressor as well as a tumor growth inducer, depending on the type of cancer. In this chapter, we review the role of SIRT6 in metabolism, genomic stability, and cancer. Further, we provide an insight into the interplay of the tumor-suppressing and oncogenic roles of SIRT6 in cancer. Additionally, we discuss the use of small-molecule SIRT6 modulators as potential therapeutics.