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. 2024 Jun 19;21:52. doi: 10.1186/s12987-024-00553-5

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© The Author(s) 2024

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/. 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. 2 — Effect of pumilio proteins on CLDN-5 expression. (a and b) The expression level of CLDN-5 mRNA (a) and CLDN-5 protein (b) in HEK-293 cells stably expressing CLDN-5 or its 3’-UTR mutants. Three independent clones were used. (c) CLDN-5 expression level was checked 48 h after the transfection of shRNA against GFP, PUM1 or PUM2 into each CLDN-5 expressing HEK-293 cells (n = 3). shRNA GFP transfected cells were used as a control. (d) FLAG-tagged N-terminally truncated PUM1 mutant expressing vectors. RD; repression domain, PCM; pumilio conserved motif, RBD; RNA-binding domain. (e) CLDN-5 expression level in HEK-293 cells stably expressing wild-type CLDN-5 was checked 48 h after the transfection of FLAG-tagged truncated PUM1 mutants expressing vectors. GFP-expressing vector was used for a transfection control. ns, not significant (p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01; ***p value ≤ 0.005;****p value ≤ 0.001