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. 2018 Jan 25;8:1570. doi: 10.1038/s41598-018-19817-w

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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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Comparison of metastasis in ache wild-type and mutant larvae. Representative metastatic larva images from Hep3B (a) and SKHep1 (b) injections into ache+/? (n = 154) larvae are shown. Green fluorescent dye DiO labeled migrated cells can be seen both in Hep3B (a’) and SKHep1 (b’) cell injections. Injection of Hep3B (c) and SKHep1 (d) into yolk sac of ache−/− (n = 51) larvae did not show a metastatic phenotype (c’,d’) (when there were more than 5 cells in tail region that embryo was counted as positive for metastasis, see (d’) which was counted as negative for metastasis). (e) Positive metastasis percent in ache wild-type and mutant groups were compared where there were significantly more metastatic larvae in ache wild-type group (P = 0.0002, chi-square test). (f) To see the effect of metastasis on local tumor size, tumor sizes of metastasis positive and negative groups were compared. Metastasis negative larvae (n = 129) showed a trend for larger tumors although non-significant (P = 0.1112, T-test) when compared to metastasis positive larvae (n = 60). n.s = not significant.