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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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CD109 levels are inversely correlated with EMT marker expression, migration, invasion in FaDu cells. (A) Isolation of CD109^H, CD109^M, and CD109^L subpopulations of FaDu SCC cells by flow cytometry based on their CD109 expression levels. (B) Representative image and (C) quantification of Western blot for EMT markers in indicated samples, showing EMT markers expressions are inversely correlated with CD109 expression. (D,F) Representative images and (E,G) qualification of immunofluorescence microscopy stained for CD109 (green), a-SMA (red, D) and Snail (Red, F) and DAPI (blue) CD109^H, or CD109^L FaDu cells, as indicated. a-SMA and Snail expressions were decreased in CD109^H FaDu cells. (H) Representative images and (I) quantification of wound healing assays. CD109 levels were inversely corelates with the motility of the FaDu cells. (J) Representative images and (K) quantification of an invasion assay. CD109 levels inversely corelated with the invasiveness of FaDu 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. Magnification, ×100. Scale bars, 100 μm.