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. 2019 Jul 25;10:3320. doi: 10.1038/s41467-019-11147-3

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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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Fig. 6 — The strength of STAT3 binding divides e4 enhancers into two subgroups. a Heatmaps of STAT3 ChIP-seq signals at cluster e4 enhancers in the four indicated conditions. Enhancers were separated into two subsets: STAT3^hie4 (n = 787) and STAT3^loe4 (n = 739) based upon a cutoff of log₂ normalized tag counts = 3). The boxplot (right) depicts normalized tag counts at STAT3^hie4 and STAT3^loe4 enhancers. b Heatmaps of STAT3 ChIP-seq signals at two subsets of e4 enhancers (defined in a) in resting and IL-10-stimulated macrophages. The boxplot (right) indicates normalized tag counts at STAT3^hie4 and STAT3^loe4 enhancers. c The most significantly enriched transcription factor (TF) motifs identified by de novo motif analysis using HOMER at STAT3^hie4 (top) and STAT3^loe4 (bottom) enhancers. d Heatmaps of CDK8 ChIP-seq signals at STAT3^hie4 enhancers in the four indicated conditions. The boxplot (right) indicates normalized tag counts at STAT3^hie4 enhancers. e Heatmaps of SMC1 ChIP-seq signals at STAT3^hie4 enhancers in the four indicated conditions. The boxplot (right) indicates normalized tag counts at STAT3^hie4 enhancers in four conditions. ****p < 0.0001, paired-samples Wilcoxon signed-rank test. f Representative UCSC Genome Browser tracks displaying normalized tag-density profiles at e4 enhancers of IL10 and TNIP3 in the indicated conditions. g Enriched Gene Ontology (GO) and MSigDB pathway categories of genes assigned to STAT3^hie4 enhancers (upper panel) or STAT3^loe4 enhancers (lower panel). h Heatmaps of IL-10-inducible cluster IV genes that correspond to STAT3^hie4-associated genes (left panel) or non-IL-10-inducible cluster IV genes that correspond to STAT3^loe4-associated genes (right panel). Data are representative of two independent experiments each of which included at least two independent donors (a, c, g, h) or depict one ChIP experiment using pooled samples from independent experiments using two (b) or four different donors (d, e). Boxes encompass the twenty-fifth to seventy-fifth percentile changes. Whiskers extend to the tenth and ninetieth percentiles. The central horizontal bar indicates the median