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. 2019 Nov 21;10(12):877. doi: 10.1038/s41419-019-2108-x

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© The Author(s) 2019

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. 2 — The TGFβ signaling pathway enhances the protein interaction between RUNX1 and p-SMAD3/ SUV39H1. a Western blotting analysis of SMAD2, p-SMAD2, SMAD3, and p-SMAD3 expression in four human glioma cell lines. Densitometric analysis of p-SMAD2 and p-SMAD3 is shown. n = 3 per group. b–e The interaction between RUNX1 and SUV39H1/SMAD3 was validated by co-IP. U251 cells overexpressing Flag-RUNX1 or Flag-SMAD3 (b) or Flag-SUV39H1 (c) in a background of TGFβ stimulation. N9 cells overexpressing Flag-RUNX1 or Flag-SMAD3 (d) or Flag-SUV39H1 (e) in the presence of LY2109761. f, g Subcellular localization of RUNX1, p-SMAD3 (f) and SUV39H1 (g) in U251 cells under normal or TGFβ protein conditions. Scale bar, 5 μm. A correlation analysis verified the positional overlap. n = 5 per group. (TGFβ #1 and TGFβ #2 indicate treatment of TGFβ protein for 4 h and 8 h. LY #1 and LY #2 indicate treatment of LY2109761 for 24 h and 48 h. ****p < 0.0001).