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. 2020 Feb 6;10:1984. doi: 10.1038/s41598-019-56877-y

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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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Neurons align their axons on FN-dependent organized Schwann cells. (a) Experimental design of Schwann cells (SC^wt, SC^Sox2/Cl2 or SC^Sox2/FnKO) co-cultured with mouse primary sensory neurons. (b) Representative confocal images of FN1 (green) and TUBB3 (white) immunostainings of primary sensory neurons co-cultured with SC^wt (129 cells), SC^Sox2/Cl2 (142 cells) or SC^Sox2/FnKO (120 cells). Nuclei were counterstained with DAPI (blue). Double head arrow shows the alignment of axons with FN fibres in SC^Sox2/Cl2. Scale bar, 100 μm. (c,d) Correlation of FN1 fibre angles of SC^wt and SC^Sox2/Cl2 with the axonal angles (TUBB3) (N = 3). (e,f) Quantification of FN1 fibres (N = 3, n ≥ 10 areas) and axonal anisotropy (N = 3, n ≥ 8 areas) in primary sensory neurons co-cultured with: SC^wt, SC^Sox2/Cl2 and SC^Sox2/FnKO. (g) Experimental design of Schwann cells (SC^wt, SC^Sox2/Cl2 or SC^Sox2/FnKO) co-cultured with human iPSC-derived motoneurons. (h) Representative confocal images of FN1 (green) and TUBB3 (white) immunostaining of human iPSC-derived motoneurons co-cultured with SC^wt (211 cells), SC^Sox2/Cl2 (231 cells) or SC^Sox2/FnKO (245 cells). Nuclei were counterstained with DAPI (blue). Double-head arrow shows the alignment of axons with FN fibres in SC^Sox2/Cl2. Scale bar, 100 μm. (i,j) Correlation of FN1 fibre angles of SC^wt and SC^Sox2/Cl2 with the angles of human iPSC-derived motoneurons (TUBB3) (N = 3). (k,l) Quantification of FN1 fibres (N = 3, n ≥ 18 areas) and axonal anisotropy (N = 3, n ≥ 9 areas) of human iPSC-derived motoneurons co-cultured with: SC^wt, SC^Sox2/Cl2 and SC^Sox2/FnKO. Ø corresponds to human iPSC-derived motoneurons cultured alone. Results are shown as the mean ± s.e.m. *p < 0.05, **p < 0.005, ***p < 0.0005.