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Chromatin interaction analysis of transcription factors and IRF8 in CD4+ T cells differentiated from wild-type mice

ABSTRACT: We report that the IRF8 protein is able to interact with chromatin during Th9 differenciation

ORGANISM(S): Mus musculus

PROVIDER: GSE99165 | GEO | 2017/10/02

SECONDARY ACCESSION(S): PRJNA387456

REPOSITORIES: GEO

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Transcriptome analysis of CD4+ T cells differentiated from wild-type and CD4cre IRF8flox mice

Project description:We report that the Th9 differenciation program is alterated in absence of IRF8 protein

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Transcriptome analysis of Th9 CD4+ T cells differentiated with or without Il-1beta

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Chromatin interaction analysis of transcription factors and NLRP3 in CD4+ T cells differentiated from wild-type and NLRP3KO mice

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Chromatin interaction analysis of transcription factors and NLRP3 in CD4+ T cells differentiated from wild-type and NLRP3KO mice

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4C-Seq identify the interaction between IRF8 promoter and rs2280381-containing region

Project description:To test if there is a physical interaction between the IRF8 promoter and the rs2280381-containing region, we conducted circular chromatin conformation capture assay in the U937 cell. For the IRF8 viewpoint group, we used CSP6I as the first digested enzyme and NlaIII as the second enzyme. For the rs2280381-containing region, we used MboI as the first digested enzyme and NlaIII as the second enzyme. We use 4C-PCR primer to construct IRF8 point view and rs2280381-containing region 4C library.

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IRF8 transcription factor controls survival and function of terminally differentiated conventional and plasmacytoid dendritic cells respectively

Project description:Interferon regulatory factor-8 (IRF8) has been proposed to be essential for development of monocytes, plasmacytoid dendritic cells (pDCs) and type 1 conventional dendritic cells (cDC1s) and remains highly expressed in differentiated DCs. Transcription factors that are required to maintain the identity of terminally differentiated cells are designated “terminal selectors”. Using BM chimeras, conditional Irf8fl/fl mice and various promotors to target Cre recombinase to different stages of monocyte and DC development, we have identified IRF8 as a terminal selector of the cDC1 lineage controlling survival. In monocytes, IRF8 was necessary during early but not late development. Complete or late deletion of IRF8 had no effect on pDC development or survival but altered their phenotype and gene expression profile leading to increased T cell stimulatory function but decreased type 1 interferon production. Thus, IRF8 differentially controls the survival and function of terminally differentiated monocytes, cDC1s and pDCs.

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4C-Seq identify the interaction between IRF8 promoter and rs2280381-containing region in human primary monocyte

Project description:To test if lncRNA AC092723.1 play a role in physical interaction between the IRF8 promoter and the rs2280381-containing region, we conducted circular chromatin conformation capture assay in the human primary monocyte. We knock down lncRNA AC092723.1 by electro-transfecting ASOs, the control group is transfected negtive ASOs with no impact on lncRNA AC092723.1.We used CSP6I as the first digested enzyme and NlaIII as the second enzyme. After constructing 4C library,we then utilize 4C-PCR primer to construct IRF8 point view 4C library.

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Project description:The role of the lineage-determining transcription factor Interferon regulatory factor 8 (IRF8) in microglia remains elucidated. Here we report the genome-wide IRF8 binding profiles in microglia at various ages. CUT&RUN methodology revealed that IRF8 starts to bind to the genome around postnatal day 9 (P9), and its number increases with age. In comparison to peritoneal macrophages, microglia IRF8 showed a cell-intrinsic binding pattern. By co-occurrence analysis, most IRF8 was H3K4me1/H3K27ac-marked enhancers, and many of them bind to the H3K27ac(high) super-enhancer regions. In IRF8KO microglia, H3K4me1 and H3K27ac profiles were altered and deposited aberrantly on the genome. This study provides a novel insight into understanding epigenetic regulation in microglia.

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Gene expression analyses in IRF8-induced monocyte differentiation

Project description:To understand the differentiation program in monocyte/macrophage differentiation, we performed ChIP-seq for IRF8 and H3K4me1 together with gene expression profiling during IRF8-induced monocyte differentiation. Both promoter-proximal and -distal binding of IRF8 associated with induction of the genes especially those related to monocytes/macrophages and immunity. DNA motif analysis for cis-regulatory elements of indirect IRF8 target genes predicted KLF4, essential for Ly6C+ monocyte development, to be a downstream transcription factor regulating the indirect target gene expression. Introduction of KLF4 into an Irf8-/- myeloid progenitor cell line induced a subset of IRF8 target genes and partially induced monocyte/macrophage differentiation. Together, this study revealed the genome-wide behavior of IRF8 and the IRF8-KLF4 axis during monocyte differentiation. Gene expressions in monocyte-like cells differentiated by IRF8 or KLF4 were measured at day 4 after retroviral transductions to myeloid progenitor cell line, Tot2. Two independent experiments were performed.

2013-01-24 | E-GEOD-38810 | biostudies-arrayexpress

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