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. 2018 Jun 27;9:2504. doi: 10.1038/s41467-018-04936-9

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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 — Smad6 is overexpressed in gliomas and predicts poor prognosis. a Smad6 expression in patient-derived glioma cells was examined by western blotting. GAPDH was used as a loading control. The relative quantification of Smad6 was listed. b Smad6 levels correlated to the proliferation capacity in patient-derived glioma cells (P- and r-values indicated, Spearman’s correlation analysis). Cells were cultured for 48 h and cell proliferation was monitored by a CCK-8 method. c Double immunofluorescence (IF) staining of Nestin and Ki67 in represented patient-derived glioma cells. Hoechst (Hoe) labeled the nuclei. Scale bars, 40 μm. d Smad6 expression correlated to tumor formation of patient-derived glioma cells (P- and r-values indicated, Spearman’s correlation analysis). Xenograft tumors derived from patient-derived glioma cells were shown in upper panel. The correlations of Smad6 to tumor volume and weight were shown in lower panel. e Immunohistochemistry (IHC) staining for Smad6 in glioma tissues. Scale bars, 50 μm. f Smad6 protein was frequently increased in nuclei of gliomas as compared it in normal brains (*P < 0.05, Unpaired t-test). g Overall survival analysis based on nuclear-Smad6 expression levels in glioma tissues. Groups were ranked according to nuclear-Smad6 IHC scores. The mean expression level of normal brains was used as a cutoff. The survival time of glioma patients with high Smad6 was significantly shorter than that of patients with low Smad6 (Log-rank χ² = 6.348, P = 0.0118)