To understand the effect of MOTS-c treatment on cast immobilization-induced skeletal muscle atrophy, we injected MOTS-c into casted mice. We then performed gene expression profiling analysis using data obtained from RNA-seq of three groups: non-immobilization group, immobilization control group, and immobilization and MOTS-c treated group. Overall design: Male C57BL/6J mice at 10 wk of age were obtained and assigned to one of the three experimental groups: non-immobilization control group (sterilized water injection for eight days, twice a day intraperitoneal injection, n = 8), immobilization control group (sterilized water injection for eight days, twice a day intraperitoneal injection, n = 8), and immobilization and MOTS-c treated group (15 mg/kg/day MOTS-c injection for 8 days, twice a day intraperitoneal injection, n = 8). Hindlimb immobilization was performed by using casting and the casting material was attached to both hinds. After eight days of hindlimb immobilization, mice were euthanized and gastrocnemius muscle was collected under anesthesia with isoflurane for RNA-seq.