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. 2014 Sep 5;6(5):1759091414548916. doi: 10.1177/1759091414548916

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© The Author(s) 2014

This article is distributed under the terms of the Creative Commons Attribution 3.0 License (http://www.creativecommons.org/licenses/by/3.0/) which permits any use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (http://www.uk.sagepub.com/aboutus/openaccess.htm).

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Figure 4. — Investigation of the spatial expression pattern of S100a4 in cultured Schwann cells and in sciatic nerves. Immunofluorescent microscopy revealed a strong cytoplasmic S100a4 expression in a subpopulation of cultured Schwann cells (a, arrow). Due to this strong immunofluorescent signal, the acquisition time was reduced, and therefore, cells still considerably expressing S100a4 appeared only weakly positive (a, asterisk). (b) In contrast to S100a4, p75^NTR signal was observed at a homogenous intensity level, indicating that those cells belong to the Schwann cell lineage. (c) Analysis of S100a4 mRNA levels revealed robustly increased expression levels in sciatic nerves derived from MAL-overexpressing mice at all investigated time points. Data were normalized to the expression of 60s, and P0 wild-type values were set to 1. The columns show the mean value of at least four experimental samples, and the error bars indicate the SD. * p ≤ 0.05. Please note that data for P0 and P5 were already presented in Figure 3. (d) Spatial expression analysis on sciatic nerves of 3-month-old MAL-overexpressing mice revealed strong S100a4 signal in Remak bundles, identified as Mbp-negative areas (d, arrows). (e) The same areas were also positive for p75^NTR, a marker for nonmyelinating Schwann cells (e, arrows). (f) Only weak expression of S100a4 was detected in wild-type sciatic nerves; its localization was comparable to MAL-overexpressing mice (f, arrows). Blue: DAPI. Bars: 20 µm.