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. 2020 Feb 13;11:869. doi: 10.1038/s41467-019-14154-6

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© The Author(s) 2020

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. 1 — a Time course of the change in paw withdrawal threshold (WD₅₀) to mechanical stimulation following PNI in contrast to sham operated rats (n = 36 rats, 18 shams and 18 PNIs). In this study, all sham or PNI rats were used between day 14 and 28 post-surgery. b Loss of IB4(+) terminals in the ipsilateral SDH (arrow). c Quantification of the number of IB4(+) terminals in sham and PNI rats (n = 82 rats, 40 shams and 42 PNIs). d Intensity profiles of IB4 and CGRP immunolabeling in the superficial ipsilateral SDH from PNI rats. e Example confocal images of gephyrin and α2 GABA_AR immunolabeling in the SDH to illustrate the binary mask segmentation of clusters (right insets). f–h Quantification of: the overall mean pixel intensity f, the mean intensity per cluster g, and the number of clusters h for selected inhibitory synaptic markers in the area defined by the loss of IB4 labeling in PNI rats and corresponding area in sham-operated rats (Fig. 1b, d). i Quantification of the number of clusters for a subset of GABA_A and glycine receptor subunits in the area defined for Fig. 1f-h. Inset: example immunolabeling in the region of interest defined by the loss of IB4 labeling. IB4, isolectin B4; CGRP, calcitonin gene related protein; VGAT, Vesicular GABA Transporter. GAD65,67, Glutamatic Acid Decarboxylases 65 and 67; WD₅₀, 50% paw withdrawal threshold. Number in each bar represents the number of sham or PNI rats used. Error bars in all panels represent S.E.M. (*P < 0.05; **P < 0.01; ***P < 0.001). Source data is available as a Source Data file.