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. 2017 Jun 9;8:15818. doi: 10.1038/ncomms15818

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Copyright © 2017, The Author(s)

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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(a) A schematic outlining the conserved sequences of the Smad3 promoter regions in Mus musculus and Homo sapiens. (b) CHIP analysis of the accumulation of HDAC1 at Smad3 promoter region in naive CD4⁺ T cells and iTreg cells. Normalized data are shown as percentage of input control (% Inp). IgG serves as a CHIP control. (c) Quantitative PCR detection of the lnc-Smad3 retrieved by HDAC1 specific antibody compared with immunoglobulin G (IgG) in the RIP assay within CD4⁺ T cells. Normalized data are shown as percentage of input control (% Inp). IgG serves as a RIP control. (d) CHIP analysis of the accumulation of HDAC1 at Smad3 promoter region in CD4⁺ T cells transduced with a control lentivirus (Lenti-CTR) or lnc-Smad3-expressing lentivirus (Lenti-lnc-Smad3) and cultured under iTreg cell-skewing conditions (with TGF-β) for 2 days. Normalized data are shown as percentage of input control (% Inp). IgG serves as a CHIP control. (e) Relative expression of HDAC1 in CD4⁺ T cells transduced and cultured as in d. Results are relative to the baseline expression of HDAC1 in unstimulated CD4⁺ T cells transduced with Lenti-CTR, set as 1. Error bars represent s.d. Student's t test. **P<0.01. NS, not significant. All data are from three independent experiments (mean±s.d. of technical triplicates).