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. 2017 Jan 31;8:14272. doi: 10.1038/ncomms14272

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Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(a) Western blots of HDAC2 and HDAC1 in lysates of crushed (Cr) and contralateral (Co) nerves of adult mice at 1 dpl (n=5), 3 dpl (n=5), 5 dpl (HDAC2, n=6; HDAC1, n=7), 12 dpl (HDAC2, n=6; HDAC1, n=8), 30 dpl (HDAC2, n=6; HDAC1, n=8) and 60 dpl (n=6), and quantification normalized to GAPDH and compared to Co=1, showing HDAC1/2 upregulation after lesion. Of note: both bands detected by the HDAC2 antibody were quantified and added together in the graph. (b) Co-immunofluorescence of HDAC2 or HDAC1 (red) with CD68 (green=macrophages) and DAPI labelling (blue=nuclei, overlay appears pink) at 12 dpl, showing upregulation in SCs (CD68-negative cells with elongated nuclei, white arrows) of crushed compared to contralateral nerves. Note that macrophages (CD68-positive cells, blue arrowheads) express variable levels of HDAC1 and HDAC2. Scale bar, 10 μm. (c) Denaturing IP of SUMO-1, HDAC2 (H2) or control Flag (ctrl) in unlesioned (no lesion) adult mouse sciatic nerve lysates, and western blot of HDAC2, showing that the ∼75 kDa band detected by the HDAC2 antibody corresponds to a SUMOylated form of HDAC2. HDAC2 and GAPDH western blots on lysates used for IP show the inputs. Representative photos of three independent experiments are shown. One-tailed (HDAC2, 12 dpl and 60 dpl; HDAC1, 5 dpl) or two-tailed Student's t-tests, unpaired (HDAC2: 3 dpl) or paired, P values: *P<0.05, **P<0.01, ***P<0.001. Values, mean; error bars, s.e.m.