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. 2020 Jul 9;10:11262. doi: 10.1038/s41598-020-67526-0

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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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mChABC increases Schwann cell migration and adhesion on aggrecan by removal of CS-GAGs. (a–f) Migration of Schwann cells on aggrecan showing (a) control, (b) LV-fGFP, (c) bChABC treated, and (d) LV-mChABC populations. Scale bar = 100 μm, arrows indicate direction of migration. Quantification of (e) cells migrating (N = 5, two-way ANOVA: cell type F(3,84) = 131.34, p < 0.0001, distance F(5,84) = 160.84, p < 0.0001) and (f) maximum distance (N = 5, one-way ANOVA: F(3,14) = 39.53, p < 0.0001). (g–k) mChABC increases cellular adhesion on aggrecan (g) to levels comparable with bChABC (N = 3, one-way ANOVA: F(3,12) = 68.61, p < 0.0001). Images showing (h) control, (i) LV-fGFP, (j) bChABC treated, and (k) LV-mChABC transduced cells. Scale bar = 100 μm. (l–p) anti-1B5 staining (red) of Schwann cells on aggrecan showing (l) control, (m) LV-fGFP, (n) bChABC treated, and (o) LV-mChABC populations immunostaned for anti-GFP (green) and Hoechst-33342 (blue) quantified in p) (N = 20, one-way ANOVA F(3,76) = 126.8, p < 0.0001). Scale bar = 40 μm. (q) Total FAK (N = 3, two-way ANOVA: cell type F(3,224) = 1.15, p = 0.329, substrate F(3,224) = 0.26, p = 0.8537) and (r) pFAK (N = 3, two-way ANOVA: cell type F(3,224) = 69.06, p < 0.0001, substrate F(3,224) = 491.82, p < 0.0001) levels in transduced cells on different substrates. Images show anti-pY397 FAK (green) staining of (s) control, (t) bChABC treated and (u) LV-mChABC transduced Schwann cells on an aggrecan substrate. Scale bar = 100 μm.