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. 2019 Aug 16;10:3716. doi: 10.1038/s41467-019-11618-7

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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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Fig. 3 — Cdh1 interacts with Src in a head-to-toe manner. a Immunoblot (IB) analysis of whole-cell lysates (WCL) and immunoprecipitates (IP) derived from 293T cells transfected with Flag-Src and HA-Cdh1 or HA-Cdc20. b IB analysis of WCL and IP derived from 293T cells transfected with HA-Cdh1 and the indicated constructs. c GST pull-down analysis to determine Src bound to purified recombinant GST-Cdh1, but not the GST recombinant protein. d Endogenous Src bound to endogenous Cdh1. IB analysis of WCL and anti-Cdh1 IP derived from MCF7 cells. e Proximity ligation assay (PLA) demonstrated a cytoplasmic interaction between Cdh1 and Src proteins in MCF7 and MDA-MB-231 cells. Scale bar, 50 μm. f IB analysis of cytoplasmic and nuclear fractions derived from the indicated cell lines. The relative band intensities of Cdh1 were quantified using ImageJ. g Schematic illustrations of the domain structures of Cdh1 and Src. h IB analysis of WCL and IP derived from 293T cells transfected with HA-Src and the indicated Flag-Cdh1 constructs. i IB analysis of WCL and IP derived from 293T cells transfected with Flag-Cdh1 WD40 domain and the indicated HA-Src constructs. j IB analysis of WCL and IP derived from 293T cells transfected with Flag-Cdh1 N terminus (aa1–174) and the indicated HA-Src constructs. k A schematic illustration of how Cdh1 interacts with Src in a head-to-toe manner. l IB analysis of WCL and IP derived from 293T cells transfected with Flag-Cdh1 WD40 domain and the indicated HA-Src constructs. m IB analysis of WCL and IP derived from 293T cells transfected with HA-Cdh1 and the indicated Flag-Src constructs