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. 2022 Jan 18;12:902. doi: 10.1038/s41598-021-04723-5

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© The Author(s) 2022

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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RFP-tagged WT and mutant variants of the Shank3 N-terminus (SPN + Ank domains), or mRFP alone, were coexpressed in 293 T cells with active (G12V-mutant) variants of the small G-proteins Rap1a (GFP-tagged; a) or HRas (HA-tagged, b). After cell lysis, RFP-tagged proteins were immunoprecipitated using the mRFP-trap matrix. Input and precipitate samples were analysed by Western blotting using epitope-specific antibodies. (c) Quantitative analysis of the data shown in (a) and (b). Signal intensities in IP samples for G-proteins were divided by IP signals for mRFP-Shank3 variants. **^,***Significantly different from WT, p < 0.01, p < 0.001; data from three independent experiments; ANOVA, followed by Dunnett’s multiple comparisons test.