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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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Neutrophils are recruited to the tumor-initiating microenvironment. Live-imaging of neutrophils in the hindbrain of Tg(mpx:mCherry) larvae expressing gfap:kras^WT or kras^G12V at 3 dpf (A). Neutrophils within the field of view were quantified and normalized to the total gfap-expressing area within the same field of view at 3 and 6 dpf (B, n = 24 kras^WT 3dpf, 42 kras^G12V 3 dpf, 24 kras^WT 6 dpf, and 30 kras^G12V 6 dpf larvae). Live time-lapse imaging was performed for 9 hours beginning at 3–3.5 dpf and total neutrophils recruited to the hindbrain during the period of imaging were quantified (C, n = 9 kras^WT and 11 kras^G12V larvae). qRT-PCR was performed on whole-larvae mRNA isolates for several neutrophil recruiting chemokines implicated in the tumor microenvironment including cxcl8a, cxcl8b, and il1b (D, n = 6 replicates for cxcl8a/b, 5 replicates for il1b, normalized as in Fig. 1D). *p < 0.05.