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. 2021 Jan 14;11:1372. doi: 10.1038/s41598-020-80621-6

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© The Author(s) 2021, corrected publication 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Validation of iL17a/f1 splicing blocking morpholino Spl-Mo and its effect on infiltration of neutrophils and macrophages in the liver. (A) Diagram of iL17a/f1 gene for the targeted sites of Spl-Mo and PCR primers. A 200-bp fragment is expected when Spl-Mo blocks the splicing. (B) Agarose gel electrophoresis of RT-PCR products after introduction of Spl-Mo. Mopholinos were introduced into zebrafish embryos at one cell stage and RNA was isolated at 6 hpf for RT-PCR analysis. (C, D) Representative images of liver-infiltrated neutrophils (C) and macrophages (D) after il17a/f1 knockdown by Spl-Mo. Lyz+ and mpeg+ transgenic zebrafish were used for marking neutrophils (dsRed expression) and macrophages (mCherry expression) respectively and these transgenic fish were compounded with kras+ zebrafish for investigation of liver-infiltrated immune cells. Livers are outlined for non-kras+ samples. (E–G) Quantification of liver size (E), number (F) and density (G) of liver-infiltrated neutrophils. (J–L) Quantification of liver size (J), number (K) and density (L) of liver-infiltrated macrophages. N = 10 per group. Scale bars: 100 μm. Statistical significance: *P˂0.05.