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. 2020 Oct 28;11:85. doi: 10.1186/s13229-020-00385-8

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© The Author(s) 2020

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Effect of Shank3 on localization of δ-catenin in neurons. a Primary rat hippocampal neurons were transfected (DIV7) with RFP-δ-catenin together with either an empty pHAGE-GFP construct as control or pHAGE-GFP-Shank3 construct. The neurons were fixed (DIV14) and stained for MAP2 as dendritic marker and PSD-95 as synaptic marker (scale bar 20 µm). In the control condition δ-catenin shows a diffused signal throughout transfected neurons, whereas coexpression of Shank3 results in recruiting more δ-catenin to postsynaptic sites with a distinct punctate pattern along dendrites. Boxed areas are magnified for GFP, RFP and PSD-95 signals (scale bar 5 μm). b Using ImageJ, targeting of overexpressed δ-catenin to the postsynaptic sites was quantified as ratio of fluorescence signal intensity in dendritic spines (positive for PSD-95) versus the signal intensity in adjacent dendritic shafts. 150 spines of 15 neurons obtained from three independent experiments per each condition were analysed. The results of quantitative analyses show that Shank3 significantly increases the level of δ-catenin in the postsynaptic sites (n = 15, unpaired T test, ****p < 0.0001, mean ± SD)