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. 2017 Jun 23;7:4202. doi: 10.1038/s41598-017-04097-7

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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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TGFβ induces Smad4 binding to SBEs in the promoter of mir-200/141 in PC9 cells. Chromatin immunoprecipitation was performed to identify whether a physical interaction between Smad4 and a predicted SBE locus in the shared promoter of mir-200c/-141 resulted from TGFβ treatment. Normal rabbit IgG served as the antibody negative control and α-Satellite primers as the negative PCR control. ID1 locus immunoprecipitation was the positive control for Smad4 binding. (a) In A549, positive Smad4-ID1 association is observed with TGFβ treatment, but an Smad4-SBE interaction is not. (b) In PC9, both Smad4-ID1 and Smad4-SBE interaction is observed. Significance was calculated using an unpaired t-test comparing TGFβ-treated cells and -untreated samples with the same primer set.