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. 2020 Feb 13;40(4):914–928. doi: 10.1161/ATVBAHA.119.313359

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© 2020 The Authors.

Arteriosclerosis, Thrombosis, and Vascular Biology is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 1. — Macrophage inflammation-suppressing transcript (Mist) is suppressed in macrophages by high-fat diet (HFD) in vivo and LPS (lipopolysaccharide) treatment in vitro. A, MA plot of log2 ratio fold change (HFD/normal diet) vs normalized mean counts. Points for differentially expressed genes (false discovery rate <0.1) are in red. B, Classification of differentially expressed genes. C and D, Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of Mist and Fabp5 in peritoneal macrophages from normal diet (ND) or HFD mice treated with or without LPS (100 ng/mL) for 3 h (n=6 mice/group). E, Representative tracks of our RNA-seq data at Mist genomic locus, and chromatin immunoprecipitation-seq tracks of overlapping histone modifications from bone marrow–derived macrophage (BMDMs; ENCODE [Encyclopedia of DNA Elements] Consortium). F, RT-qPCR analysis of genes expressed in adipose tissue macrophages (ATMs) from mice fed ND or HFD for 16 wk (n=4 mice/group). G, Nuclear localization of Mist relative to cytosol in macrophages. Subcellular fractionation of RAW 264.7 cells followed by RT-qPCR for indicated genes, including Neat1, a known nuclear lncRNA. H and I, RNA-fluorescent in situ hybridization in mouse peritoneal macrophages and RAW cells for Mist (yellow foci, denoted by white arrows); nuclei are counterstained with DAPI (blue). Scale bar represents 10 μm. Bar graphs represent mean±SD. H3K27ac indicates lysine-27 acetylation; and H3K4me1, H3 lysine-4 monomethylation. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 calculated using Fisher least significant difference test on log-transformed expression values.