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. 2018 Mar 14;9:1075. doi: 10.1038/s41467-018-03425-3

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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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Fig. 1 — Laminin-α1 and laminin-α5 deposition in the niche of activated satellite cells. a Quantitative PCR analysis of Lama1-5, Lamb1-3, and Lamc1-3 genes together with Pax7 and MyoD in non-injured (dark gray; n = 3) and 4 dpi injured (light gray; n = 3) TA muscles. b Schematic representation of the myofiber culture system showing the sequential activation (blue), proliferation (yellow), self-renewal (green), and differentiation (purple) of satellite cells. c Representative immunofluorescence images of cultured myofibers analyzed for laminin-α1 (green), Pax7 (red), Myf5 (yellow at 24 h), and MyoD (yellow at 72 h). Panels on the right are high magnification images of the area boxed. White arrows indicate satellite cells associated with laminin-α1. Scale bar, 50 μm. d Immunofluorescence analysis of laminin-α5 (green) and Pax7 (red) distribution in cultured myofibers. Arrows indicate Laminin-α5 deposition in Pax7⁻cells at 72 h. Scale bar, 50 μm. e Quantification of the number of satellite cells expressing Pax7, MyoD, and laminin-α1 after myofiber culture. The color code relates to the cell populations described in (b). n = 3 experiments with 20–32 fibers analyzed per time point. Graph shows mean + sem; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (t-test)