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. 2019 Dec 20;9:19560. doi: 10.1038/s41598-019-56163-x

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© The Author(s) 2019

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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Fluorescence microscopy of the rat right gastrocnemius muscle, 2 days after turpentine oil injection. (a) Rhodamine labeled D-P8RI positive signal (black) is concentrated at the border of the turpentine oil injection. (b) Higher magnification in the blue inset reveals that D-P8RI labeled cells are enriched at the immediate vicinity of the damaged tissue. (c) Immunofluorescence of another rat right gastrocnemius muscle after turpentine oil injection. (d) The inner layer of the cells concentrated around the site of turpentine oil injection is mainly composed of CD68+ leucocytes (Fuchsia). (e) The mononuclear and polynuclear leukocytes (identified by the DAPI staining) at the immediate vicinity of the damaged tissue (red inset) display a cleavage of CD31 as revealed by the relative negative extracellular CD31 (eCD31) signal (green) in spite of a consistent intracellular CD31 (iCD31) staining (red). (f) The cleavage of CD31 was instead rather limited at the surface of the leukocytes located at the periphery of the inflammatory site (green inset). Of note, in parallel with a lesser CD31 cleavage phenomenon, the D-P8RI signal appears weaker in such an outer layer of the inflammatory site, as documented in (b).