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. 2017 Oct 24;7:13859. doi: 10.1038/s41598-017-14313-z

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© The Author(s) 2017

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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The quantity of SMN protein in exosomes reflects the intracellular levels in fibroblasts derived from a mouse model of SMA and patients with type I SMA. Panel A: Crude protein cell lysates (5 µg) from MEFs derived from wildtype mice (MEF^WT) or Smn ^2B/− mice (MEF^2B/−), a mouse model of SMA, were analysed by immunoblot for SMN and tubulin (loading control). Data is representative of n = 3. Panel B: Exosomes were isolated from the medium of MEF^WT and MEF^2B/−, 3 µg of the resulting samples were separated by SDS-PAGE and analysed by immunoblot for SMN, Alix, and flotillin (loading control). Data is representative of n = 3. Panel C: Crude protein cell lysates (5 µg) from normal, SMA carrier or SMA type I human fibroblasts were analysed by immunoblot for SMN and tubulin (loading control). Data is representative of n = 3, and is quantified in Panel D. Asterisks (*) indicates significant differences between groups, determined by Bonferroni post-hoc analysis (p < 0.05). Panel E: Exosomes were isolated from the medium of normal, carrier or SMA type I fibroblasts, 3 µg of the resulting samples were separated by SDS-PAGE and analysed by immunoblot for SMN, Alix, and flotillin (loading controls). Data is representative of n = 3, and is quantified in Panel F. Asterisks (*) indicates significant differences between groups, determined by Bonferroni post-hoc analysis (p < 0.05).