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Cerebral dopamine neurotrophic factor (CDNF) is an endoplasmic reticulum (ER) resident protein, and its expression and secretion are induced when tissues and cells are subjected to hypoxia, ischemia, or traumatic injury. As a novel cardiomyokine, CDNF plays a crucial role in the progression of myocardial I\/R injury. In our previous study, we reported that the overexpression of CDNF inhibited tunicamycin\u2010induced H9C2 cell apoptosis. Moreover, there is a unique <jats:italic>N<\/jats:italic>\u2010glycosylation site at Asn57 in the CDNF protein, which likely affects its function in H9C2 cells. However, the detailed impact remains unexplored. In our current study, we observed elevated levels of CDNF in the serum of acute MI patients, myocardial tissue of I\/R model mice, and H\/R model H9C2 cells. To detect the effect of <jats:italic>N<\/jats:italic>\u2010glycosylation on the CDNF protein, we constructed an Asn57 mutant (N57A) plasmid and found that the N57A protein presented similar intracellular localization to those of the wild\u2010type CDNF protein. However, the N57A protein demonstrated reduced stability, and the mutant protein could not protect H\/R\u2010induced H9C2 cells from apoptosis. Moreover, this process may occur through the downregulation of the PI3K\/Akt pathway. Therefore, <jats:italic>N<\/jats:italic>\u2010glycosylation of CDNF may be essential for protein stability and its protective role in H\/R injury in H9C2 cells.<\/jats:p>","DOI":"10.1002\/cbin.70000","type":"journal-article","created":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T07:12:11Z","timestamp":1739257931000},"page":"472-483","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Regulation of <i>N<\/i>\u2010Glycosylation of CDNF on Its Protein Stability and Function in Hypoxia\/Reoxygenation Model of H9C2 Cells"],"prefix":"10.1002","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-9470-6861","authenticated-orcid":false,"given":"Qingwen","family":"Huang","sequence":"first","affiliation":[{"name":"Qingdao Medical College Qingdao University Qingdao Shandong China"},{"name":"Department of Cardiology Yantai Yuhuangding Hospital Yantai Shandong 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