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. 2019 Nov 21;9:17276. doi: 10.1038/s41598-019-53660-x

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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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Representative images of CTCs isolated by DEPArray. The upper panel shows spiking experiments performed by using MDA-MB231 and MCF-7 BC cell lines predominantly expressing mesenchymal (M: N-cadherin/CD146/CD44; PE, red) and epithelial markers (E: E-cadherin/EPCAM; FITC, green), respectively. The lower part of the figure shows four different CTC sub-populations from the same blood sample (patient #242), showing variable expression of M and/or E markers as well as negative blood and endothelial markers (CD45/CD31/CD34; APC, purple). Nuclei are stained by Hoechst 33342 (blue). The first line shows a CTC expressing only M markers (M+/E−). Line 2 represents a CTC lacking both E and M markers (M−/E−). The APC fluorescence detectable near M−/E− CTC was due to non-nucleated blood components, namely erythrocytes and platelets that in several instances bind CTCs. Line 3 shows a CTC expressing both M and E markers (M+/E+), whereas line four depicts a CTC with E phenotype (M−/E+). Two lymphocytes derived from the CD45+ fraction of the same blood sample, are also shown as control for blood cell markers (purple), with or without CD44 expression (red).