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Epigenetic events regulate muscle mass and function maintenance. We hypothesized that the pattern of epigenetic events (muscle\u2010enriched microRNAs and histone acetylation) and acetylation of transcription factors known to signal muscle wasting may differ between early\u2010 and late\u2010time points in skeletal muscles of mice exposed to hindlimb immobilization (I) and recovery following I. Body and muscle weights, grip strength, muscle\u2010enriched microRNAs, histone deacetylases (HDACs), acetylation of proteins, histones, and transcription factors (TF), myogenic TF factors, and muscle phenotype were assessed in gastrocnemius of mice exposed to periods (1, 2, 3, 7, 15, and 30 days, I groups) of hindlimb immobilization, and in those exposed to reloading for different periods of time (1, 3, 7, 15, and 30 days, R groups) following 7\u2010day immobilization. Compared to non\u2010immobilized controls, muscle weight, limb strength, microRNAs, especially miR\u2010486, SIRT1 levels, and slow\u2010 and fast\u2010twitch cross\u2010sectional areas were decreased in mice of I groups, whereas Pax7 and acetylated FoxO1 and FoxO3 levels were increased. Muscle reloading following splint removal improved muscle mass loss, strength, and fiber atrophy, by increasing microRNAs, particularly miR\u2010486, and SIRT1 content, while decreasing acetylated FoxO1 and FoxO3 levels. In this mouse model of disuse muscle atrophy, muscle\u2010enriched microRNAs, especially miR\u2010486, through Pax7 regulation delayed muscle cell differentiation following unloading of gastrocnemius muscle. Acetylation of FoxO1 and 3 seemed to drive muscle mass loss and atrophy, while deacetylation of these factors through SIRT1 would enable the muscle fibers to regenerate. J. Cell. 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