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. 2017 Dec 13;8:2116. doi: 10.1038/s41467-017-02029-7

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© The Author(s) 2017

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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Fig. 7 — OTUD1 inhibits lung metastasis in transplantable mouse models of breast cancer metastasis. a–d Control (Co.) or OTUD1-silenced (sh#1 and sh#2) 4T07 cells were tail vein-injected into nude mice. Lung metastasis was measured by BLI. Normalized photon flux at the indicated times left (±SD) and representative images (right) are shown; p-values; Student’s t-test (a). Lung metastasis nodules in each group were calculated (±SD) and the representative images of bright view (c, upper panel; scale bar, 2 mm) and bioluminescent view (c, lower panel; scale bar, 2 mm) are shown. Representative HE and IHC staining pictures of lung sections of control and one shRNA-depleted OTUD1 group (d), scale bars, 1 mm and 20 μm for lung haematoxylin and eosin (H&E) and SMAD7 immunohistochemistry images, respectively). e–i Control (Co.) 4T1 cells or cells stably expressing OTUD1-wt or CA were tail vein-injected into nude mice. Lung metastasis was measured by BLI. Normalized photon flux (±SD) at the indicated times are shown in the upper panel and representative images in the lower; p values; Student’s t-test e. Lung metastasis nodules (±SD) f and average lung lesion surface area (±SD) g (arbitrary units based on pixel quantification from digital images) in each group were calculated and the representative images of bioluminescent view (h, scale bar, 2 mm) are shown. Representative HE and IHC staining pictures of lung sections of every experimental group (i, Scale bars, 1 mm and 20 μm for lung haematoxylin and eosin (H&E) and SMAD7 immunohistochemistry images, respectively) are shown. j–m Control (Co.) or OTUD1-depleted 4T1 cells were orthotropic injected in syngeneic BALB/c mice. Primary tumor formation in mice after orthotropic injection of each group at day 21 after implantation. Normalized BLI signals (±SD) (Left) and representative BLI signals of primary tumor. Number of lung metastasis nodules (k) and average lung lesion surface area (±SD) (l) (arbitrary units based on pixel quantification from digital images). Representative bioluminescent view of isolated lung (m, left, scale bar, 2 mm) and HE staining of lung section of each group (m, right, scale bar, 20 μm). *p < 0.05 and **p < 0.01 (two-tailed Student′s t-test a–l)