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. 2020 Aug 14;10:13841. doi: 10.1038/s41598-020-70800-w

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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 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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EnMT investigation upon bFGF and TGF-β on rCEnC. (a) The panel shows representative immunofluorescence images of α-SMA (red, first row) and S100A4 (green, second row) in rCEnC treated with Mock control, bFGF, TGF-β and bFGF + TGF-β, respectively. White arrows indicate the cells positive for α-SMA. In blue DAPI, scale bar 50 µM for all the images. (b) The bar chart on the right shows the percentage of cells positive for α-SMA and the percentage of cells in which S100A4 moved to the nuclei as a mean of 12 fields (n = 3 biological replicates) for each condition. Results are presented as mean ± SE. T-test was performed n.s. non-significant. (c) The panel illustrates a representative image of a double immunostaining with S100A4 in green, DAPI in blue and α-SMA in red of rCEnC at a high passage number (P10). Letters P, perinuclear, and N, nuclear, underlie the different localization of S100A4 staining, corresponding to a low and high α-SMA positivity, respectively. (d) The panel shows a secondary only control on Mock rCEnC, used as a negative control with DAPI in blue.