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. 2020 Feb 13;9(2):16. doi: 10.1038/s41389-020-0204-5

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© The Author(s) 2020

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. 4 — a Immunoblot for TG2 in control, wild-type (WT), and TGM2-deleted (KO) 4T1 cells. Expression of β-tubulin (β-tub) was used as a loading control. b Control (WT) and TGM2-deleted (KO) 4T1 cells were seeded under single-cell 3D culture conditions. Initiation of 3D outgrowth was quantified by bioluminescence. Data are normalized to the plated values and are the mean ± SD of three independent analyses resulting in the indicated p value. (below) Representative brightfield images of each 3D culture. c Control (WT) and TGM2-deleted (KO) 4T1 cells were engrafted onto the mammary fat pad and primary tumor growth was quantified by caliper measurements. Data are of individual mice taken at the indicated time points, resulting in the indicated p value. d Quantification of bioluminescent radiance from the pulmonary regions of interest (ROIs) at the indicated time points. (below) Representative thoracic bioluminescent images of control 4T1 (WT) TG2-deleted (KO) tumor-bearing mice. e Upon necropsy, the numbers of metastatic pulmonary nodules was quantified from control (WT) and TGM2-deleted (KO) 4T1 tumor-bearing mice. (below) Representative gross anatomical images of lungs from these groups. f Upon necropsy, the lungs and primary tumors of mice bearing control (WT) and TGM2-deleted (KO) 4T1 tumors were removed, and the carcasses were immediately imaged to visualize extra pulmonary metastases (arrows). A representative mouse from each group is shown. For d, e, data are the mean ± SE of five mice resulting in the indicated p values.