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. 2018 Sep 5;9(9):906. doi: 10.1038/s41419-018-0902-5

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© The Author(s) 2018

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. 4 — a Dual-luciferase assay results of Twist1, Twist2, SNAIL1, and SNAIL2. PPI inhibited the Twist1 promoter activity in a dose-dependent manner. b Immunofluorescence results showed that PPI inhibited Twist1 expression in the nucleus. c Western blot results showed that induced hypoxia upregulated Twist1 and VM markers; this effect was reversed by PPI. d PPI also enhanced Twist1 downregulation in SMMC7721 cells. The expression levels of Twist1, vimentin, VEGFR1, VEGFR2, and VE-cadherin decreased, whereas that of E-cadherin increased. e Dual-luciferase assay of E-cadherin, VE-cadherin, VEGFR1, and VEGFR2. PPI decreased the expression levels of VE-cadherin, VEGFR1, and VEGFR2 and increased that of E-cadherin. f ChIP-PCR assay showed that PPI inhibited the binding of Twist1 to the promoter of VE-cadherin. The values are means of three experiments, and error bars represent standard deviation (n = 3, *P < 0.05, **P < 0.01)