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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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Primary hippocampal neurons expressing GFP-Shank3 (WT or P141A, L270M mutants) were fixed and stained for the postsynaptic marker PSD-95 and the dendritic marker MAP2 (gray). The expressed Shank3 variants were detected using the GFP fluorescence. (a) Overview images of neurons (scale bar 20 μm). (b) Boxed dendritic areas were magnified (scale bar 10 μm). GFP-Shank3 colocalises extensively with PSD-95 in presumably postsynaptic clusters, which are found in a small distance from the (MAP2-positive) main dendrite. (c) Quantitative evaluation of the total number of Shank3 clusters per 20 μm dendrite. ns: non-significant; analysis of 45 dendrites of n = 15 neurons from three independent experiments; t-Test. (d) Quantitative evaluation of the number of postsynaptic Shank3 clusters (Shank3 clusters colocalising with PSD-95) per 20 µm dendrite. *Significantly different from WT, p < 0.05; analysis of 45 dendrites of n = 15 neurons from three independent experiments; t-Test. (e) Quantitative evaluation of the number of primary dendrites per neuron. ns: non-significant; analysis of n = 15 neurons from three independent experiments; t-Test.