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. 2018 Jan 19;9(2):59. doi: 10.1038/s41419-017-0090-8

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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. 8 — a Schematic representation of human SMAD6 promoter reporter constructs. Fragments of various lengths between −2155 and −1540 bp of SMAD6 were cloned downstream of the firefly luciferase reporter. b Luciferase activity in HEK293T cells transfected with firefly luciferase reporter plasmids containing various upstream regions of SMAD6. A Renilla luciferase reporter was cotransfected with a pGL3-basic or plasmid reporter. c Fragments of various lengths between −1961 and −1730 bp of SMAD6 were cloned downstream of the firefly luciferase reporter. d Luciferase activity in HEK293T cells transfected with firefly luciferase reporter plasmids containing various upstream regions of SMAD6. e Putative transcription factor binding sites in the region between −1860 and −1730 bp upstream of SMAD6 (upper panel). Luciferase activity associated with the region between −1860 and −1730 bp of SMAD6 in HEK293T cells transfected with small interfering RNA (siRNA) against HIC2, NR4A2 or with a negative control (NC) (lower panel). f HIC and SMAD6 expression in SK-Hep 1 and Huh-7 cells after HIC2 was knocked down as assessed with real-time PCR (upper panel). NR4A2 and SMAD6 expression in SK-Hep 1 and Huh-7 cells after NR4A22 was knocked down (lower panel). Data represent the mean ± SEM. **P < 0.01; ***P < 0.001