Project description:The transcription factor IRF8 is a critical regulator of plasmacytoid dendritic cell (pDC) and classical dendritic cell (cDC) development in both mouse and man. Yet the downstream molecular targets that regulate DC homeostasis and development are largely unknown. A recent study using gene expression analysis of IRF8-deficient myeloid and lymphoid progenitors identified the Myc paralog Mycl1 as a potential transcriptional target of IRF8. We report here that Mycl1 is a mediator of DC homeostasis at steady state and during inflammation, and its expression is regulated by IRF8 in multiple DC lineages. We have further validated these observations with ChIP-Seq of IRF8 binding to the Mycl1 locus. Notably, IRF8 binding to Mycl1 locus is independent of an interaction with the AP1 factor, BATF3. Additionally, our genome-wide survey of IRF8 binding identified both EICE and AICE motifs. Examination of IRF8 binding in dendritic cells
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.
Project description:To understand the mechanism underlying monocyte and dendritic cell development through the regulation of Irf8 expression by the 56 kb downstream (+56 kb) Irf8 enhancer, we performed epigenetic profiling of bone marrow cells and splenocytes from wild-type, the Irf8 +56 kb enhancer-deficient, and IRF8-deficient mice. Taken together with the transcriptome analysis of mononuclear phagocyte lineage cells in these mice, the Irf8 +56 kb enhancer-mediated high Irf8 expression in hematopoietic progenitor cells promote type 1 classical dendritic cell (cDC1) differentiation, while low Irf8 expression in progenitors led to Ly6C+ monocyte development. In addition, IRF8 ChIP-seq of mature cDC1s and monocytes suggested that IRF8 regulates enhancers in cooperation with different transcription factors in each lineage in its expression level.
Project description:To understand the effect of BCR-ABL and IRF8 on gene expression pattern during in vitro dendritic cell differentiation, microarry analysis was performed. Murine bone marrow Lin- cells were transduced with retroviruses carrying human BCR-ABL cDNA and/or mouse Irf8 cDNA, and then cultured with Flt3L for 5 days. Two independent experiments were performed.
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.
Project description:To understand the effect of BCR-ABL and IRF8 on gene expression pattern during in vitro dendritic cell differentiation, microarry analysis was performed.
