Project description:We have performed analyses of murine primary bone marrow derived monocytes challenged with either PBS or oxLDL. oxLDL selectively enhances growth and adhesion potential of monocytes. Purified bone marrow monocytes were treated with PBS or 20 microgram/ml oxLDL for a five day period, and cells were harvested for scRNAseq analyses.
Project description:Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of lipid-loaded macrophages in the arterial wall. Intimal macrophages internalize modified lipoproteins such as oxidized LDL (oxLDL) through scavenger receptors, leading to storage of excess cholesteryl esters in lipid bodies and a "foam cell" phenotype. In addition, stimulation of macrophage Toll-like receptors (TLRs) has been shown to promote lipid body proliferation. We investigated the possibility that there are transcriptional regulators that are common to both pathways for stimulating foam cell formation (modified lipoproteins and TLR stimulation), and identified the transcription factor ATF3 as a candidate regulator. In this specific microarray experiment, we studied the effect of genetic knockout of ATF3 on the transcriptional response of macrophages to oxLDL. Murine bone marrow-derived macrophages from two different mouse strains (Atf3-/- and WT) were incubated in the presence or absence of oxidized low-density lipoprotein (oxLDL), and then transcriptionally profiled using the Affymetrix Mouse Exon Array 1.0 ST. The goal was to study the pattern of differential expression between WT and Atf3-/- strains, in oxLDL-induced foam cells and in non-foamy control cells, to identify cellular pathways that may be dysregulated under loss of the transcription factor ATF3 in macrophage foam cells. Twelve female mice (six Atf3-/-, and six WT control mice) were sacrificed at 8-12 weeks of age, and macrophages were derived from the femoral bone marrow using rhM-CSF. oxLDL was introduced into the medium for six samples (3 WT and 3 Atf3-/-) at 25 ug/mL on day seven, and after 24 h of incubation, RNA was isolated using Trizol. Labeled cRNA derived from the RNA samples was hybridized to Affymetrix Mouse Exon Array 1.0 ST GeneChips. Microarray data were processed using transcript-level probesets, and are in log2 scale.
Project description:Bone marrow derived macrophages of Lysz-Cre;Catnbtm2Kem(fl/fl) mouse were compared with bone marrow derived macrophage of Catnbtm2Kem(fl/fl) control mouse Total RNA extracted from bone marrow derived macrophage
Project description:To get insight into TRIM33 functions, TRIM33 ChIP-seq was carried out in murine macrophage cell line (RAW) and in bone marrow-derived macrophages (BMDM). The results showed that, in addition to its role in hematopoietic differentiation, TRIM33 may modulate PU.1 transcriptional activity during macrophage development and/or activation.To characterize the role of TRIM33 in macrophages, we bred TRIM33fl/fl mice with Lyz-Cre mice where the Cre recombinase gene is under the regulatory sequences of the Lyz gene that is expressed only in mature myeloid cells. Bone marrow cells from LyzCre/Trim33+/+ mice and LyzCre/Trim33flox/flox mice were differentiated in macrophages and treated during 0h, 4h, 12h and 24h with LPS. Using ChIP-seq, we provide a link between TRIM33 binding and H3K4me3 spreading on inflammatory genes in macrophages. Chromatin immunoprecipitations of TRIM33 and H3K4Me3 followed by multiparallel sequencing performed in murine bone marrow-derived macrophages (BMDM).
Project description:to determine whether hydroxymethyl butyrate alters macrophage polarization bone marrow derived macrophages were treated with HMB alone or in combination with LPS for 48h
Project description:1) Differential gene expression profiling of bone marrow derived macrophage treated with GapmeR NS (control) or MERRICAL (KD) in both M0 and M1 phase
Project description:Transcriptional profiling of Aza+ITF-2357 treated bone marrow derived macrophages vs. Mock treated bone marrow derived macrophages. Aim was to elucidate the effects of epigentic drug treatment on in vitro cultured macrophages and to compare these data to in vivo treated macrophage populations. Transcriptional profiling of NSCLC cell lines treated with epigenetic agents Transcriptional profiling of immune populations in the lung Kras G12D tumor micronevironment treated with combination epigenetic therapy Transcriptional profiling of lung Kras G12D tumors treated with combination epigenetic therapy
