Stitutes of Health, Bethesda, USA) and normalized to -actin24. The uncropped scan with the blot is reported in the Supplementary Fig. 7. Reside Pralidoxime Activator animal imaging. Macrophage localization in vivo was obtained by NIR imaging with the fluorescent label macrophage mice by utilizing PhotonImager (Biospace Laboratory, Paris, France)83. Mouse thioglycollate-elicited peritoneal macrophages had been harvested (up to 250 106 cells per ml) and incubated for 15 min at RT with VivoTrack 680 (PerkinElmer, Inc., Waltham, USA), dissolved in sterile PBS, washed, centrifuged (400 g, ten min) and diluted to a final concentration of five 106 cells 40 l-1. Retro-orbital vein injection (40 l) of labeled macrophages was performed in pSNLsham C57BL6 mice at day 9 after surgery. Twenty-four h later,NATURE COMMUNICATIONS | eight:| DOI: 10.1038s41467-017-01739-2 | www.nature.comnaturecommunicationsNATURE COMMUNICATIONS | DOI: 10.1038s41467-017-01739-ARTICLE21. Taylor-Clark, T. E., Ghatta, S., Bettner, W. Undem, B. J. Nitrooleic acid, an endogenous product of nitrative tension, activates nociceptive sensory nerves through the direct activation of TRPA1. Mol. Pharmacol. 75, 82029 (2009). 22. Trevisani, M. et al. 4-Hydroxynonenal, an endogenous aldehyde, causes discomfort and neurogenic inflammation through activation of the irritant receptor TRPA1. Proc. Natl Acad. Sci. USA 104, 135193524 (2007). 23. Bautista, D. M. et al. TRPA1 mediates the inflammatory actions of environmental irritants and proalgesic agents. Cell 124, 1269282 (2006). 24. Trevisan, G. et al. Novel therapeutic technique to stop chemotherapy-induced persistent sensory neuropathy by TRPA1 blockade. Cancer Res. 73, 3120131 (2013). 25. Obata, K. et al. TRPA1 induced in sensory neurons contributes to cold hyperalgesia right after inflammation and nerve injury. J. Clin. Invest. 115, 2393401 (2005). 26. Katsura, H. et al. Antisense knock down of TRPA1, but not TRPM8, alleviates cold hyperalgesia just after spinal nerve ligation in rats. Exp. Neurol. 200, 11223 (2006). 27. Caspani, O., Zurborg, S., Labuz, D. Heppenstall, P. A. The contribution of TRPM8 and TRPA1 channels to cold allodynia and neuropathic pain. PLoS One particular four, e7383 (2009). 28. Eid, S. R. et al. HC-030031, a TRPA1 selective antagonist, attenuates inflammatory- and neuropathy-induced mechanical hypersensitivity. Mol. Pain four, 48 (2008). 29. McNamara, C. R. et al. TRPA1 mediates formalin-induced pain. Proc. Natl Acad. Sci. USA 104, 135253530 (2007). 30. Trevisan, G. et al. TRPA1 mediates trigeminal neuropathic discomfort in mice downstream of monocytesmacrophages and oxidative strain. Brain 139, 1361377 (2016). 31. Kim, H. K. et al. Reactive oxygen species (ROS) play a vital part within a rat model of neuropathic discomfort. Pain 111, 11624 (2004). 32. Liu, T., van Rooijen, N. Tracey, D. J. Depletion of macrophages reduces axonal degeneration and hyperalgesia following nerve injury. Discomfort 86, 252 (2000). 33. Zhu, X., Fujita, M., Snyder, L. A. Okada, H. Systemic delivery of neutralizing antibody targeting CCL2 for glioma therapy. J. Neurooncol. 104, 832 (2011). 34. Perkins, N. M. Tracey, D. J. Hyperalgesia Formic acid (ammonium salt) Cancer because of nerve injury: part of neutrophils. Neuroscience 101, 74557 (2000). 35. Hackel, D. et al. The connection of monocytes and reactive oxygen species in pain. PLoS A single eight, 2013 (2013). 36. Szolcsanyi, J., Szallasi, A., Szallasi, Z., Joo, F. Blumberg, P. M. Resiniferatoxin: an ultrapotent selective modulator of capsaicin-sensitive key afferent neurons. J. Pharmacol. Exp. Ther. 255, 923.