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. 2016 May 12;14:133. doi: 10.1186/s12967-016-0876-y

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© Malara et al. 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 4 — Phenotypic characterization of eCTCs. a Calpain expression was compared in the same patient between CTCs (collected before surgical treatment) and correspondent tumour tissue sample collected during surgical resection. Control α-tubulin immunoblot confirmed equal loading. b Bar graph displayed eCTCs dynamic phenotypic changes in different stage of tumour (according to Tumor-Node-Metastasis, UICC-2009) (data are presented as mean ± standard deviation). Multiple comparison procedures pairwise showed significant differences between advanced (T3N1M0, T4N1M1) versus localized colon cancer stages (T1N0M0, T2N0M0). *p < 0.05, **p ≤ 0.01. c–f, immunofluorescence characterization of eCTCs for epithelial (Epcam, green channel) and mesenchymal (Fibronectin, red channel) antigens expressions. Their co-expression on cell’s surface revealed the EMT phenotype (merged color image). Scale bars 50 μm