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. 2020 Jul 6;39:128. doi: 10.1186/s13046-020-01638-3

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© The Author(s) 2020

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. 4 — NCSTN induces EMT in HCC cells via activation of Zeb1. a The mRNA expression levels of EMT markers CDH1, CDH2, VIM, ZEB1, SNAIL1, SNAIL2, FOXC1, FOXC2 and TWIST1 in the indicated HCC cells. b Luciferase reporter assays showed relative luciferase activity of Zeb1 promoter in indicated cells. c The expression levels of four EMT-related proteins E-cadherin, Zeb1, Vimentin and N-cadherin in the indicated cells. Loading control was assessed by β-actin. d Immunofluorescence assays showed the expression of NCSTN, E-cadherin, Vimentin and Zeb1 in the indicated cells. Scale bars, 100 μm. e The expression levels of EMT-related markers E-cadherin, Vimentin and N-cadherin in the indicated cells co-transfected with siZeb1 or negative control. Loading control was assessed by β-actin. f The migration and invasion capacity was examined in the indicated HCC cells co-transfected with siZeb1 or negative control. g Wound healing assays showed the migration capacity of indicated HCC cells. h Representative images of EMT-related markers E-cadherin, Vimentin and N-cadherin in HCC tissues by immunohistochemical staining. Scale bar, 100 μm. i Correlation analyses of protein expression levels between NCSTN and E-cadherin, Vimentin or nuclear Zeb1 in HCC tissues. *p < 0.05, **p < 0.01, ***p < 0.001