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. 2022 Jun 9;13:839677. doi: 10.3389/fimmu.2022.839677

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Copyright © 2022 Wu, Quan, Zhang, Wang, Ou, Mao, Sun, Yang, Wu, Wei, Jia, Chen, Zhu, Liu, Zhao, Zhang, Huang, Gao, Tian and Cheng

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Attenuated DHAV maintains high m6A levels in duckling liver. (A) Quantification of m6A modification in total RNAs and mRNAs from duckling livers using m6A antibody-based immunoblotting. Methylene blue staining served as the loading control. Representative dot blot images at 12 hpi and 36 hpi are displayed. m6A-modified mRNA is present at greater levels in the CH60-infected group compared with the CH-infected group (middle panel). (B) A more sensitive LC–MS/MS approach was used to further quantify m6A RNA (m6A/A) and m6A mRNA (m6Am/A) in duckling livers infected by a virulent CH strain and attenuated CH60 strain. The virulent CH strain significantly reduces m6A modification levels in the poly-A tailed mRNA pool. However, m6Am (an m6A-resembling modification) modification levels remain similarly low in both groups. Data are displayed as the mean ± SD with 4 replicates.