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. 2018 Jul 19;8:10909. doi: 10.1038/s41598-018-29101-6

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© The Author(s) 2018

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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Effects of NMac1 on in vitro invasion/migration of MDA-MB-231 cells and in vivo metastasis. (a) NMac1 inhibits transwell migration and matrigel invasion of MDA-MB-231 cells. MDA-MB-231 cells treated with indicated concentrations of NMac1 were allowed to migrate in Boyden chambers for 24 hours. Bars indicate percentage of migration and invasion. ^*P < 0.05, Student’s t-test. (b) NMac1 inhibits matrigel invasion of MDA-MB-231 cells via Nm23-H1 and H2. Nm23-H1 or (/and) H2 knocking down MDA-MB-231 cells treated with 25 µM of NMac1 were allowed to invaded in matrigel coated Boyden chambers for 24 hours. Bars indicate amount of migration and invasion. ^*P < 0.05, Student’s t-test. (c) NMac1 inhibits transwell migration of MDA-MB-231 cells via Nm23-H1 and H2. Nm23-H1 or (/and) H2 knocking down MDA-MB-231 cells treated with indicated concentrations of NMac1 were allowed to migrated in Boyden chambers for 24 hours. Bars indicate percentage of migration and invasion. ^*P < 0.05, Student’s t-test. (d) MDA-MB-231-Luc-D3H2LN cells were orthotopically injected into NOD/SCID mice and the mice were treated with vehicle or NMac1 (n = 7 for VEH group, n = 6 for NMac1 group). (Left) Bioluminescence images visualizing the metastatic tumor cells. (Right) Quantification of bioluminescence images 3 weeks after the NMac1 treatment. Data are presented as mean total photon flux per second ± S.D.