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. 2023 Oct 14;13:17447. doi: 10.1038/s41598-023-44731-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Expression of H antigens in the COS-7 cells transfected with various FUT1 constructs. The FUT1 allele containing uncharacterized SNP(s), wild-type allele (positive control), or FUT1*01N.09 (with c.725T>G) subcloned into pcDNA3.1(+) plasmids was transfected into COS-7 cells. Negative control is COS-7 cells transfected with pcDNA3.1(+) plasmid without FUT1 allele. After 2 days of culture, the cells were incubated with 1E3 mouse monoclonal antibody to H type 1–4, followed by incubation with FITC-conjugated goat anti-mouse IgM secondary antibody, and expression of H antigen on the cell surface was monitored by flow cytometry. Nine representative flow cytometry results including positive and negative controls are shown.