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. 2017 Nov 21;7:15918. doi: 10.1038/s41598-017-15139-5

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© The Author(s) 2017

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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Enhancement of the EMT in CRC cells by USP47 overexpression under hypoxic conditions. (a) DLD-1 cells were transfected with the pcDNA3-USP47 construct and then incubated under normoxic conditions for 72 h. Expression levels of USP47, E-cadherin, N-cadherin, and vimentin were evaluated by Western blotting. (b) The morphology of mock- and pcDNA3-USP47-transfected DLD-1 cells was visualized by phase contrast microscopy. Scale bar = 20 μm. (c) Immunocytochemical analysis of pcDNA3-USP47-transfected DLD-1 cells stained for USP47, E-cadherin, N-cadherin, and vimentin under normoxic conditions. Scale bar = 20 μm. (d) Representative images of colony formation in pcDNA3-USP47-transfected DLD-1 cells. Scale bar = 200 μm. (e) Representative images of enhanced wound closure of pcDNA3-USP47-transfected DLD-1 cells subjected to hypoxic conditions. Scale bar = 100 μm. Data in (d,e) are representative of three independent experiments. Two-sided student’s t-tests were performed to assess significance; *P < 0.05, **P < 0.01, ***P < 0.001, and ^# P < 0.01.