DNA methylation dynamics of Flt3L induced ex vivo BMDC differentiation
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ABSTRACT: To understand the DNA methylation dynamics during Flt3L induced Bone-marrow derived dendritic cell (BMDC) differentatiion we performed reduced representation bisulfite sequencing (RRBS). Three distinct stages were chosen and included the initial bone marrow cells, the Common Myeloid Progenitor (CMP), and fully differentiated BMDC.
Project description:To understand the gene expression dynamics during Flt3L induced Bone-marrow derived dendritic cell (BMDC) differentatiion we performed RNA-seq. Three distinct stages were chosen and included the initial bone marrow cells, the Common Myeloid Progenitor (CMP), and fully differentiated BMDC.
Project description:DNGR-1 (CLEC9A) is expressed in CD8-like dendritic cells (DCs) and detects a ligand exposed on necrotic cells. We have characterized the transcripts that are induced in cultures of Flt3L-derived bone-marrow-derived WT or DNGR1-deficient DCs upon incubation with dead cells. Microarray analysis of the transcriptome of DNGR-1-deficient and WT DCs cultured with dead cells failed to reveal any DNGR-1-dependence in the transcriptional response to dead cells. Flt3L-BMDC from WT or DNGR-1-deficient (Clec9a gfp/gfp) mice were left untreated or were cultured for 5 hours with UV-treated dead cells at a 1:2 ratio. CD24hi CD11blow B220- CD8α+-like DC were purified by cell sorting and RNA was extracted. There were 3 replicates for each of the four experimental conditions.
Project description:To assess genotypic differences between IL-33-induced CD103+ cDC1s and GM-CSF-induced CD103+ cDC1s IL-33 or GM-CSF was treated at 5 ng/ml on the day 5 of Flt3L-BMDC generation and then the cells were incubated for an additional 5 days Then, we performed RNA sequencing of CD103+ cDC1s isolated from IL-33 or GM-CSF-treated Flt3L-BMDCs
Project description:Analysis of baseline gene expression in bone marrow derived dendritic cells (BMDC) from female CBA/J (CBA) and C57BL/6 (BL/6) mice. Results provide insight into strain-dependent differences in gene expression.
Project description:Analysis of baseline gene expression in bone marrow derived dendritic cells (BMDC) from female CBA/J (CBA) and C57BL/6 (BL/6) mice. Results provide insight into strain-dependent differences in gene expression. CBA and BL/6 BMDCs prepared from individual mice were plated in replicate at 1x10^6 cells/ml in 48-well tissue culture plates (BD Falcon). Replicates were pooled after 4hr and total RNA was obtained by TrizolM-BM-. (Invitrogen) extraction according to the manufacturerM-bM-^@M-^Ys instructions.
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.