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. 2022 Apr 2;21:93. doi: 10.1186/s12943-022-01537-5

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 8 — CircMAP3K4-455aa was ubiquitinated by MIB1 and degraded by the ubiquitin–proteasome pathway. A Representative confocal images showing the cytoplasmic location of circMAP3K4-455aa. B The half-life of circMAP3K4-455aa protein after treatment with cycloheximide. C Western blot assay examining circMAP3K4-455aa expression levels after treatment with MG132, NH₄Cl, or 3-Methyladenine. D, E. Co-IP (D) and confocal colocalization (E) verified circMAP3K4-455aa interaction with MIB1. F Western blot detection of circMAP3K4-455aa after MIB1 knockdown (left panel) and MIB overexpression (right panel). G In vivo ubiquitylation assay assessing circMAP3K4-455aa ubiquitination after MIB1 knockdown. H Detection of the circMAP3K4 half-life after decreasing MIB1 expression. CHX, Cycloheximide; cMAP, circMAP3K4; 3-MA, 3-Methyladenine; IP, Immunoprecipitation