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Therefore, our study aimed to examine whether <jats:italic>O<\/jats:italic>\u2010GlcNAcylation affected myogenic IL\u20106 expression, regulation of energy metabolism, and promotion of survival in mouse skeletal muscle under acute cold exposure conditions. Total protein was extracted from C2C12 cells that had been cultured at 32\u00b0C for 3, 6, 9, and 12\u2009h. Western blot analysis showed that mild hypothermia enhanced <jats:italic>O<\/jats:italic>\u2010GlcNAc transferase (OGT) and <jats:italic>O<\/jats:italic>\u2010GlcNAcase (OGA) expression. Furthermore, global OGT\u2010dependent glycosylation and interleukin\u20106 (IL\u20106) levels peaked 3\u2009h after induction of mild hypothermia. Enhanced activation of the NF\u2010\u03baB pathway was also observed in response to mild hypothermia. Alloxan and Thiamet G were used to reduce and increase global OGT glycosylation levels in C2C12 cells, respectively. Increased <jats:italic>O<\/jats:italic>\u2010GlcNAcylation was associated with significant upregulation of IL\u20106 expression, as well as enhanced activity and nuclear translocation of p65, while decreased <jats:italic>O<\/jats:italic>\u2010GlcNAcylation had the opposite effect. In addition, increased <jats:italic>O<\/jats:italic>\u2010GlcNAcylation was associated with significantly increased glucose metabolism, and OGT\u2010mediated <jats:italic>O<\/jats:italic>\u2010GlcNAcylation of p65. We generated skeletal muscle\u2010specific OGT knockout mice and exposed them to cold at 4\u00b0C for 3\u2009h per day for 1 week. OGT deficiency attenuated the <jats:italic>O<\/jats:italic>\u2010GlcNAcylation, activity, and nuclear translocation of p65, resulting in downregulation of IL\u20106 in mouse skeletal muscle of mice exposed to cold conditions. Taken together, our data suggested that <jats:italic>O<\/jats:italic>\u2010GlcNAcylation of p65 enhanced p65 activity and nuclear translocation leading to the upregulation of IL\u20106, which maintained energy homeostasis and promotes cell survival in mouse skeletal muscle during cold exposure.<\/jats:p>","DOI":"10.1002\/jcp.30612","type":"journal-article","created":{"date-parts":[[2021,10,21]],"date-time":"2021-10-21T11:44:43Z","timestamp":1634816683000},"page":"1341-1352","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["OGT upregulates myogenic IL\u20106 by mediating <i>O<\/i>\u2010GlcNAcylation of p65 in mouse skeletal muscle under cold exposure"],"prefix":"10.1002","volume":"237","author":[{"given":"Yajie","family":"Hu","sequence":"first","affiliation":[{"name":"National Experimental Teaching Demonstration Center of Animal Medicine Foundation, College of Animal Science and Veterinary Medicine Heilongjiang Bayi 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