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. 2018 Jun 15;8:9168. doi: 10.1038/s41598-018-27601-z

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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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Interaction between SLIT2 and ROBO1 and ROBO2 as determined by the coimmunoprecipitation experiment. The chicken GCs were transfected with the pYr-adshuttle-4-SLIT2 expression construct. After transfection for 24 h, the cells were lysed, and the lysates were immunoprecipitated with a control chicken IgG or an antibody against HA. The immunoprecipitation (IP) with the HA antibody was analyzed by immunoblotting (IB) using the SLIT2, ROBO1 and ROBO2 antibodies. After the lysates were immunoprecipitated with the chicken IgG, no pYr-adshuttle-4-SLIT2 band was observed in the immunoprecipitants from the cells expressing HA-SLIT2, and no ROBO1 and ROBO2 bands were observed (IP: chicken IgG). After the lysates were immunoprecipitated with an antibody against HA, endogenous ROBO1 and ROBO2 were coimmunoprecipitated in the cells expressing HA-SLIT2 (IP: HA). All blots were cropped, and the gels were run under the same experimental conditions.