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. 2021 Mar 3;11:5085. doi: 10.1038/s41598-021-84106-y

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© The Author(s) 2021

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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LOTUS is distributed in the synapse region. (a) LOTUS expression in cultured hippocampal neurons (DIV14). The image was acquired using a confocal microscope. PSD-95 (blue), LOTUS (red), and Bassoon (green). Scale bar, 10 µm. (b) Magnified image from (a). The segment was imaged at 5 × magnification. Scale bar, 5 µm. (c) Super-resolution image of Bassoon (green) and LOTUS (red) at the synapse site. The image was acquired using a STED microscope. Scale bar, 1 µm. (d) Super-resolution image of PSD-95 (green) and LOTUS (red) at the synapse site. The image was acquired using a STED microscope. Scale bar, 1 µm. (e) LOTUS and NgR1 localization in cultured hippocampal neurons (DIV14). The image was acquired using a confocal microscope. PSD-95 (blue), LOTUS (red), and NgR1 (green). Scale bar, 10 µm. (f) Magnified image from (e). The segment was imaged at 3 × magnification. Scale bar, 10 µm. (g) Quantification of LOTUS expression against that of Bassoon, PSD-95 and/or NgR1. Bars indicate ratio of the colocalization of PSD-95 and LOTUS in PSD-95 positive puncta (PSD95-LOTUS/PSD95), that of Bassoon and LOTUS in Bassoon positive puncta (BSN-LOTUS/BSN), that of NgR1, LOTUS and PSD-95 in PSD-95 positive puncta (NgR1-LOTUS-PSD95/PSD95) and that of NgR1, LOTUS and PSD-95 in PSD-95 and NgR1 positive puncta (NgR1-LOTUS-PSD95/PSD95-NgR1), respectively. Data are means ± SEM from 3–14 cells. **P < 0.01, one-way ANOVA post-hoc Tukey–Kramer.