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. 2018 Sep 5;8:13285. doi: 10.1038/s41598-018-31675-0

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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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Blocking neutrophil motility reduces proliferation of tumor-initiating cells. Wildtype or Tg(mpx:mCherry) embryos were co-injected with gfap:GFP-kras^G12V plasmid and rac2 or standard control morpholino (MO). Larvae were live-imaged for neutrophil recruitment at 3 dpf (A) and quantified for neutrophils within the field of view normalized to GFP-expression area (B, n = 13 control MO and 18 rac2 MO larvae). Larvae were also fixed at 3 dpf and immunostained for phospho-Histone H3 (C). Double-positive (pH3 and GFP) cells were quantified by optical sectioning and normalized as above (D, yellow arrowheads in C, n = 21 control MO and 17 rac2 MO larvae). Tg(mpx:mCherry-2a-Rac2^WT) or Tg(mpx:mCherry-2a-Rac2^D57N) embryos were injected with gfap:GFP-kras^G12V and live-imaged at 3 dpf (E). mCherry-positive neutrophils were quantified and normalized as above (F, n = 52 Rac2^WT and 46 Rac2^D57N larvae). Larvae were fixed at 3 dpf and immunostained for phospho-Histone H3 (G). pH3-GFP double positive cells were quantified by optical section and normalized as above (H, n = 30 Rac2^WT and 37 Rac2^D57N larvae). *p < 0.05.