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. 2019 Nov 6;9:16317. doi: 10.1038/s41598-019-50694-z

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© The Author(s) 2019

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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rhCD109 rescue the EMT phenotype of CD109-KO SCC cells. (A) Phase contrast microscopy for the morphology of parental A431 cells (Left), CD109-KO A431 cells (Middle), and CD109-KO cells treated with rhCD109 protein (Right), demonstrated that rhCD109 rescued the epithelial phenotype of CD109-KO cells. (B) Western blot detection and (C) quantification of E-Cadherin (CHD1), fibronectin (FN), alpha-smooth muscle actin (a-SMA), Snail and Twist. The treatment of rhCD109 protein rescue E-cadherin expression and significantly suppressed the expressions of mesenchymal proteins and EMT markers. (D–F) Representative Immunofluorescence microscopy images and (G) quantification for CDH1 (green, D), a-SMA (red, E), Snail (red, F) and DAPI (blue) in parental A431 cells, CD109 KO cells, and CD109-KO cells treated with rhCD109 as indicated, confirming that the treatment of CD109 suppressed EMT markers and rescued the epithelial traits in CD109-KO cells. All the results are expressed as the mean ± S.D. of three independent experiments. Significance is calculated using a One-Way ANOVA *P < 0.05. **P < 0.01 and ***P < 0.001. Scale bars: 20,30 and 200 μm in low and high magnification, as indicated.