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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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Cxcr1 promotes neutrophil recruitment to the tumor-initiating microenvironment. Adult cxcr1 heterozygote zebrafish expressing Tg(mpx:mCherry) were in-crossed, injected with gfap:kras^G12V, imaged for neutrophil recruitment to the hindbrain at 3 dpf, and then processed for single-embryo genotyping (A). Neutrophils recruited to the field of view were quantified in cxcr1 mutants and WT or heterozygote siblings (no difference observed between +/+ and +/−) and normalized to total gfap-expressing area (B, n = 40 WT/het and 44 cxcr1 mutant larvae). Tg(mpx:mCherry) larvae expressing gfap:kras^WT or kras^G12V were treated beginning at 2.5 dpf with 1 µM SB225002 or 0.02% DMSO vehicle control for 16 hours followed by live-imaging (C). Neutrophil recruitment was quantified and normalized as above (D, n = 42 DMSO and 43 SB225002 larvae). Wildtype Tg(mpx:mCherry) embryos were co-injected with gfap:GFP-kras^WT or kras^G12V and standard control MO or cxcr1 splice-blocking morpholino. Larvae were live-imaged at 3 dpf (E) and neutrophil recruitment was analyzed as described previously (F, n = 12 kras^WT, 21 kras^G12V ctrl MO, and 26 kras^G12V cxcr1 MO larvae). *p < 0.05, One-way ANOVA with post-hoc Tukey’s test performed for data in 4 F.