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. 2018 Sep 11;8:13638. doi: 10.1038/s41598-018-31835-2

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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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Identification of phospho-STAT3 binding sites in fatty dHepaRG by ChIP-seq analysis. dHepaRG cells were treated with sodium oleate 250 μM for 4 days (Oleate) or vehicle treated (Ctrl) and immunoprecipitated with a phospho-Tyr705-STAT3 specific antibody. (a) Candidate phospho-STAT3 binding sites from MACS2 peak calling (default MACS2 parameters, qvalue < 0.01). (b) Motif enrichment analysis: STAT (signal transducer and activator of transcription), IRF (interferon-regulatory factor). (c/d) Chip-seq profile (left panels) showing phospho-STAT3 enrichment (black circles) on miR-21 (c) and ISG15 (d) promoters after sodium oleate 250 μM treatment. Right panels: cross-linked chromatin from dHepaRG cells treated as in (a) was immunoprecipitated with a phosphoTyr705-STAT3 (pSTAT3) and an acetylated-Histone4 (AcH4) antibody and analyzed by qPCR with primers specific for the observated phospho-STAT3 peaks on miR-21 (c) and ISG15 (d) promoters, identified by chipseq analysis as shown in left panels. Histograms show Fold Induction (FI) of the % of Input (mean from 3 independent experiments; bars indicate S.D.; asterisks indicate p-value).