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. 2018 Jan 17;8:964. doi: 10.1038/s41598-018-19301-5

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© The Author(s) 2018

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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P2RX4 expression in Dorsal Root Ganglion neurons. (A) Representative cropped western blotting analysis of lumbar DRG extract indicates the presence of a 60 kDa band in P2RX4^+/+ DRG that is absent in extracts from P2RX4^−/− DRG. N = 3 independent experiments, n = 3 mice per genotype. (B) β-galactosidase staining was used as a surrogate marker for P2RX4 in DRG sections. Only P2RX4^−/− neurons are immuno-positive for ßgal compared to sections from P2RX4^+/+ DRG. Scale bar 100 µm. (C) DRG neurons that express ß-gal innervate paw skin. Fluoro-Gold B, a retrograde marker of neurons was injected in paw-skin and L5/L6 DRG were removed for staining a week later. βgal immunostaining reveals a population of neurons that are labeled for both ßgal and Fluoro-Gold B. Scale bar 50 µm. (D) In nociceptive neurons, ßgal is mainly expressed in c-ret-, TRPV1- and IB4-positive neurons, but scarcely in others populations (CGRP, TrkA or Substance P (SP) expressing cells). Representative images of DRG sections stained with βgal and makers of nociceptive neurons. Scale bars 50 µm.