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. 2019 Feb 6;9:1509. doi: 10.1038/s41598-018-36605-8

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© The Author(s) 2019

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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Expression of cancer angiogenesis genes in TANs and effects of neutrophil depletion on liver tumorigenesis and angiogenesis. (A) Heatmap representation of expression of cancer angiogenesis genes extracted from DEGs of TANs. The color codes represent fold changes of expression of each gene in each sample relative to the mean expression of all samples. The list of cancer angiogenesis genes was obtained based on IPA knowledge datbase. (B) Representative confocal images of 6-dpf lyz+ larvae injected with different morpholinos: MO_SC (left, n = 18) or MO_gcsfr (right, n = 20). (C) Numbers of neutrophils in the middle body after injection of morpholinos. Neutrophils were counted in the the middle body surrounding the liver region (from the posterior edge of eye to posterior edge of swimbladder). (D) Representative 3D confocal images of 6-dpf kras−/fli+ and kras+/fli+ larvae injected with MO_SC or MO_gcsfr. For fli+ larvae, both Tg(fli1a::GFP) (left) and Tg(fli1a::RFP) (middle and right) were used. Kras- larvae were LiPan strain (left) with DsRed expression in the liver. (C and D) Quantification of blood vessel density in the liver area (E) and liver size (F) after morpholino injection. N >20 in each group. Statistical significance: *p < 0.05.