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Here, we have carried out a systematic study to find the mechanisms of NO\u2010induced regulation of RBC deformability under hypoxia.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>NO levels, RBCs membrane elongation index (EI), membrane protein band 3 methemoglobin (MetHb) were determined during hypoxia (0 to 120\u2009minutes). To validate the role of NO in regulating RBC deformability, tests were also performed with a NO donor (sodium nitroprusside) or a NO synthase inhibitor (<jats:sc>l<\/jats:sc>\u2010nitro\u2010arginine methylester) under 60\u2009minutes hypoxia.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Hypoxia for 45\u2009minutes increased NO levels from 25.65\u2009\u00b1\u20091.95 to 35.26\u2009\u00b1\u20092.01\u2009\u03bcmol\/L, and there was a plateau after 60\u2009minutes hypoxia. The EI did not change before 45\u2009minutes hypoxia, but decreased from 0.567\u2009\u00b1\u20090.019 to 0.409\u2009\u00b1\u20090.042 (30\u2009Pa) after 60\u2009minutes hypoxia. The cross\u2010linking of band 3 and phosphotyrosine increased after 45\u2009minutes hypoxia. All can be alleviated by supplement NO and aggregated by inhibiting NOS. However, the MetHb was not present this trend.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>NO may prevent decreased of RBCs deformability through reducing the cross\u2010linking of membrane band 3 under hypoxia; this helps microvascular perfusion of RBCs during ischemic and hypoxic disease states.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcb.27359","type":"journal-article","created":{"date-parts":[[2018,9,16]],"date-time":"2018-09-16T12:54:22Z","timestamp":1537102462000},"page":"305-320","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Nitric oxide inhibits hypoxia\u2010induced impairment of human RBC deformability through reducing the cross\u2010linking of membrane protein band 3"],"prefix":"10.1002","volume":"120","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3751-3892","authenticated-orcid":false,"given":"Yajin","family":"Zhao","sequence":"first","affiliation":[{"name":"Key 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