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. 2021 Mar 4;11:5232. doi: 10.1038/s41598-021-84667-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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Multiplex in situ hybridization shows co-expression of eleven neuropeptides. Five combinations of three neuropeptides each: (A–C) Tac1, Cck and Nts; (D–F) Nms, Nmu and Tac2; (G–I) Npff, Grp and Nmb; (J–L) Trh, Grp and Tac1; (M–O): Tac1, Ucn3 and Tac2 in green, red and blue, respectively. (A,D,G,J,M) Overview images over the lumbar dorsal horn. The outline of the gray matter is indicated. Scale bar 100 μm. Box indicates position of the higher magnification image in (B,E,H,K,N). (C,F,I,L,O) Number of cells per animal (two hemi-sections) that (co-)express the three neuropeptides. Circle and intersection sizes are approximately proportional to the cell number they represent.