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. 2018 May 8;8:7262. doi: 10.1038/s41598-018-25435-3

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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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Sema3A binds Domain IV-3 and shRNA against Sema3A abrogates C4-2 cell clustering. (A) Domain IV-3 was adsorbed to well surfaces and subjected to a binding assay with Sema3A-Fc, Sema3A-Fc and Sema3A antibody, Sema3A antibody alone, and control IgG. Sema3A-Fc binds to Domain IV-3 and Sema3A antibody diminishes the binding signal significantly to 250 ng/mL (student’s unpaired t-test per concentration). (B) Blossom62 alignment of Ig 17 of perlecan (2^nd of Domain IV-3) and the Ig of Sema3A. C4-2 cells were seeded on Domain IV-3 after being transduced with either control shRNA (panel C) or Sema3A directed shRNA (panel D). (E) Dispersion index quantification between control and directed shRNA for each substrate type and amount. Significance values are student’s unpaired t-test between control and Sema3A shRNA. p-values: *<0.05, **<0.01, ***<0.001.