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. 2017 Jun 19;8:15870. doi: 10.1038/ncomms15870

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Copyright © 2017, The Author(s)

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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(a) A549-luc-Vector and A549-luc-miR-128-3p cells of the indicated dosages were, respectively, implanted in the inguinal folds of different nude mice. Representative bioluminescent images of subcutaneous tumour outgrowth are shown. (b) Subcutaneous tumours formed by the indicated cells were dissected and imaged. H&E histologically confirmed tumour cells. Scale bar, 25 μm. (c and d) For the spontaneous metastasis model, bioluminescent images of subcutaneous tumours of the indicated cells, distant metastasis signals (with tumours shielded), and ex vivo organ metastases are shown. (e) For the experimental metastasis model, bioluminescent images of systemic metastases and ex vivo organ metastases including those in the lungs, liver, spleen, kidney, colon, heart, stomach, bones and brain, are shown. (f) Immunostaining for the lung adenocarcinoma marker mucin 1 (MUC1) and lung squamous cell carcinoma marker cytokeratin 5 (CK5), respectively, in spontaneous and experimental lung metastatic lesions developed by subcutaneous inoculation (s.c.) and intravenous injection (i.v.) of the indicated cells. Scale bar, 25 μm. (g) Immunostaining of two key EMT biomarkers, E-cadherin and Vimentin, in primary subcutaneous tumour tissues and lung metastatic lesions. Scale bar, 25 μm. H&E, haematoxylin and eosin.