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. 2019 Mar 29;9:5372. doi: 10.1038/s41598-019-41870-2

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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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Higher SMAD4 but not SMAD2 levels increase the fraction of cells with a sustained ctgf response. (A) Single cell ctgf responses after stimulation with different doses of TGF-β, clustered into transient and sustained classes using k-means clustering (n transient = 1666; n sustained = 427). Heat maps for ctgf traces belonging to transient (upper panel) and sustained (lower panel) are shown using absolute (left) and normalized levels of ctgf responses. (B) Silhouette plot of cells sorted according to ctgf expression dynamics. (C) Fraction of cells responding in a transient or sustained manner at different concentrations of dox in the iS2 (left) and iS4 (right) cell lines. (*p < 0.001, **p < 10e-6; Chi-squared test). (D) Average SMAD4/2 expression level during the 12 hours following 5 nM TGF-β stimulation, belonging to the transient and the sustained classes from samples treated with 2 ng/ml and 10 ng/ml doxycycline (*p < 0.05; t-test).