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. 2018 Jun 27;9:2504. doi: 10.1038/s41467-018-04936-9

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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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Fig. 6 — MH2 domain is essential for the interaction of Smad6 with PIAS3. a Smad6 bound to PIAS3. HA-PIAS3 was expressed in 293T cells with or without Flag-tagged nuclear-Smad6 construct (Flag-Smad6-NLS), followed by IP with anti-Flag antibody. b PIAS3 interacted with Smad6. HA-PIAS3 was expressed in 293T cells with or without Flag-Smad6-NLS, followed by IP with anti-HA antibody. c Schematic diagram of Smad6 and its deletion mutants. d Mapping of the domain of Smad6 for its interaction with PIAS3. 293T cells were co-transfected with HA-PIAS3 and Flag-tagged nuclear-Smad6 deletion mutants, followed by IP with anti-HA antibody and IB analysis with indicated antibodies. e MH2 of Smad6 could bind to PIAS3. 293T cells were co-transfected with HA-PIAS3 and His-MH2-NLS or Flag-Smad6-NLS and followed with an IP analysis using the indicated antibodies