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. 2021 Sep 17;4:1097. doi: 10.1038/s42003-021-02628-7

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Schematic of the rat SNVP splitting into four-terminal branches prior to entering into the spleen. Inserts: Splenic vasculature (yellow arrowheads) and the SN (red arrowheads). b Toluidine blue (TB) staining, red arrows point to the location of the SN fascicles between the artery (A) and vein (V). c unmyelinated SN axons by TEM with a single myelinated axon (blue arrow). d Representative images of TB staining of the four-terminal SN branches, with nerve fascicles in inserts (red arrowheads). e TEM micrographs of axonal composition in each terminal branch. Myelinated axons (blue arrows) were observed in SN1–2, but not in SN 3–4. f High TEM magnification of SN-2 axons. g Axon diameter distribution in the SN branches. h Linear regression analysis of axon diameter distribution. Kolmogorov–Smirnov normality test was used to determine the difference in axon diameter population in different groups (n = 4 animals). Data are shown as mean (solid lines) and error (95 confidence interval; top for SN1 and SN4, and bottom for SN3 and SN2). SN1–4 are shown by different colors. SN splenic nerve, SNVP splenic neurovascular plexus, TEM transmission electron microscopy. The slope between SN-1 and SN-4 is significantly different (p < 0.03).