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. 2021 May 26;11:11057. doi: 10.1038/s41598-021-90736-z

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© The Author(s) 2021

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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The I324M mutant have no obvious influence on V2R protein’s production and stability compared with WT. (A) HEK293T cells transiently expressing either WT-V2R (WT) or I324M-V2R (I324M), 48 h after transfection, cells were incubated with 20 μg/ml cycloheximide (CHX) for 0 ,4 ,8 and 12 h before detected by western blotting using anti-Myc rabbit polyclonal antibody. (B) Density of degraded protein bands were normalized against endogenous HSP90 and quantified by ImageJ 1.49 (https://imagej.nih.gov/ij/) The data was from four independent experiments.