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. 2023 Sep 8;13:14790. doi: 10.1038/s41598-023-42046-9

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© The Author(s) 2023

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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Non-overlapping expression of PRGamide and EAAT1 protein in the N. vectensis nervous system. (a) Immunofluorescence co-staining using anti-EAAT1 antibody (Bio-Techne) and anti-PRGamide antibody during the primary polyp stage (7dpf). EAAT1 staining and PRGamide staining do not overlap. EAAT1-positive cells lack processes, unlike PRGamide-positive neurons, which extend neurites (arrows). (b) PRGamide-expressing neurons detected at different developmental stages. Side views of (from left to right): early planula stage (4dpf), planula stage (5dpf), juvenile polyp stage (7dpf). Asterisks indicate oral positions. Large and small scale bars are 100 μm and 10 μm, respectively.