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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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Single cell variability of SMAD4/2 translocation dynamics and ctgf expression. (A–D) and (I–L) Heatmaps of single cell SMAD4/2 translocation (A,B,I,J) and ctgf expression (C,D,K,L) upon 5 nM TGF-β stimulation. Data is represented both in absolute luminescence levels (A,C and I,K) and intensities normalized on the maximal intensity for each trace (B,D,J,L). (E,M) Population average of SMAD4/2 translocation and ctgf expression. Vertical dashed lines: timing of the stimulation (black), SMAD translocation peak (green) and ctgf expression response initiation (blue). The latter two time points are manually annotated in single cell traces and averages of the population are indicated with dashed lines. (F,N) Single cell distribution of peak timing for SMAD4/2 translocation (green) and ctgf expression (blue). SMAD4: Mean = 55 min, CV = 0.33; SMAD2: Mean = 66 min, CV = 0.35. (G,O) Single-cell distribution of nuclear to cytoplasmic ratio at translocation peak for SMAD4 (G) and SMAD2 (O). (H,P) Single-cell distribution of ctgf expression peak levels for iS4 (H) and iS2 (P) cell lines.