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. 2020 May 5;39(23):4538–4550. doi: 10.1038/s41388-020-1307-3

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© The Author(s) 2020

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a The predicted target sequence of miR-1258 in 3′UTR of Smad2 (Smad2-3′UTR) and Smad3 (Smad3-3′UTR) and mutant containing three altered nucleotides in the seed sequence of miR-1258 (miR-1258-mut). b Luciferase assay of pGL3-Smad2-3′UTR and pGL3-Smad3-3′UTR in the presence of miR-1258 mimics, miR-1258-mutant, and anti-miR-1258. c The Smad2/3 mRNA levels after transfected by miR-1258 mimics, miR-1258-mutant, and anti-miR-1258. d The protein levels of P-Smad2, Smad2, P-Smad3, Smad3, PAI-1, E-cadherin, and vimentin after transfected by miR-1258 mimics and anti-miR-1258 in HCC cell. e The correlation between Smad2/3 gene expression and miR-1258 level in TCGA dataset. f The correlation between Smad2/3 gene expression and miR-1258 level in SYSUCC dataset. g, h The wound-healing assay and transwell assay of HCC cells transfected with miR-1258, miR-1258+Smad2, miR-1258+Smad3. Experiments performed in triplications. miR-1258, HCC cells were transfected by miR-1258 expression vector. Anti-miR-1258, HCC cells were transfected by miR-1258 antisense plasmid. miR-1258+Smad2, co-overexpressed miR-1258, and Samd2 in HCC cells. miR-1258+Smad3, co-overexpressed miR-1258, and Samd3 in HCC cells. *P < 0.05.