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. 2022 Sep 23;20:426. doi: 10.1186/s12967-022-03633-y

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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. 5 — USP4 deubiquitination maintains ACE2 stability and thus inhibits EMT and fibrosis in LUAD. A Analysis of USP4 expression in LUAD and normal lung tissues using the GEPIA database. B Analysis of USP4 expression in relation to OS of LUAD patients using Kaplan–Meier plotter. C Detection of USP4 mRNA expression in BEAS-2B and A549 by RT-qPCR. D, E Detection of USP4 protein expression in BEAS-2B and A549 by WB and statistical quantification by ImageJ software. F RT-qPCR detection of USP4 mRNA expression levels after overexpression of USP4. G, H WB detection of USP4 protein expression levels after overexpression of USP4 and statistical quantification by ImageJ software. I Overexpression of USP4 followed by RT-qPCR for EMT and fibrosis marker expression. J, K The protein expressions of TGF-β1, E-cadherin, vimentin, ACTA2 and COL1A1 were detected by WB after overexpression of USP4. Statistical quantification by ImageJ software. L, M After overexpression of USP4, cell migration was monitored by wound scratch assay. N, O After the expression of USP4, cell invasion was determined by invasion assay. And counted statistically by ImageJ. P, Q After overexpression of USP4, an immunofluorescence assay was performed to detect the expression of fibrosis-associated proteins. Statistical data are shown. R Detection of ACE2 mRNA expression by RT-qPCR after overexpression of USP4. S, T Detection of ACE2 protein expression by WB after overexpression of USP4 and quantified by ImageJ software statistics. U Functional enrichment analysis of USP4 by Metascap website. V Analysis of ubiquitinated degradation of ACE2 after overexpression of USP4 by Co-IP assay