Project description:We performed ribosome profiling and RNA-seq on mouse bone marrow derived macrophages (BMDMs) treated with 100ng/ml LPS for 0hr, 1hr, 2hrs, 4hrs, 6hrs to obtain global mRNA translational landscapes during this inflammatory response. Overall design: Examine transcriptomic and translatomic dynamics in mouse BMDMs' response to LPS treatment. Ribo-seq, RNA-Seq, CLIP-seq.
Project description:mRNA microarray analysis of bone marrow derived macrophages treated under four conditions, including Naïve (N). Bone marrow derived macrophages (BMDM) were derived from the bone marrow of mice and cultured in the presence of PAO, IFN-gamma, or lipopolysaccharide (LPS). Profiled groups include Naive, LPS, IFN, PAO. Compared each of the groups (PAO, LPS, IFN) with Naïve group.
Project description:Lipopolysaccharide (LPS)/immunue complex (IC) stimulated macrophages produce cytokine profiles that differe from LPS-stimulated inflammatory macropahges. Notch signaling is activated in LPS/IC-stimulated macrophages and inhibition of Notch signaling reduced IL-10 production. This study investigated the effect of gamma-secretase inhibitor (GSI) that suppresses Notch signaling pathway, on gene expression profiles in macrophages activated by LPS/IC. Overall design: Gamma secretase inhibitor (GSI)-treated bone marrow derived macrophage mRNA profiles were generated by deep sequencing using Illumina NextSeq 500. The results were compared between vehicle control (dimethylsulfoxide)-treated cells and GSI-treated cells.
Project description:To investigate how the phenotype of macrophages that have engulfed engineered nanoparticles (ENPs) differs from normal macrophages, we conducted Affymetrix microarray studies to identify the gene regulatory pathways affected by the ENPs. To mimic potential occupational exposure scenarios, the experimental design involved pretreatment of mouse primary bone marrow macrophages with the ENPs (25 mg/ml) for 24 hr, followed by removal of residual ENPs and challenging the macrophages with the TLR4 ligand and surrogate bacterial stimulus, lipopolysachharide (LPS) for 4 hr. The 4 hr challenge time was chosen based on preliminary studies which showed many of the proinflammatory gene expression responses peak between 2-6 hr after LPS treatment. Transcriptional responses were measured by global microarray analysis of mouse primary bone marrow macrophage cells. Groups (N=3 replicates) of primary mouse bone marrow macrophages were pretreated with selected engineered nanoparticless (33 nm iron oxide(SPIO) or 50 nm amorphous silica) at concentrations of 25 mg/ml for 24 hours. After removing the nanoparticles, macrophages were challenged with fresh media containing 10ng/ml lipopolysaccharide (LPS).
Project description:Purpose: The goal of this study is to compare downstream genes of Sema6D signaling in LPS plus IFNg stimulated macrophages. Methods: Bone marrow derived macrophage mRNA profiles of 7 weeks of wild type (WT) and Sema6D-/- mice were stimulated by LPS for 4 hrs. Results: According to this comparison, we found that 550 genes were downregulated in Sema6D-/- macrophages than WT macrophages in response to LPS. Conclusions: Our study represents 62 genes were supressed in both M1 and M2 Sema6D-/- macrophage than WT macrophages, suggesting of Sema6D reverse sigaling genes. Overall design: Bone marrow derived macrophage mRNA profiles of 7 weeks of wild type (WT) and Sema6D-/- mice were stimulated by LPS for 4 hrs, then isolated total RNA by RNeasy kit.
Project description:We report the genome-wide RNA sequencing analysis in Il10-/- bone marrow-derived macrophages (BMDMs) stimulated by lipopolysaccharide (LPS) where IL-10 effect in macrophage inflammatory response was examined in IL-10-deficient BMDMs upon LPS stimulation with addition of exogenous IL-10. Overall design: Examination of IL-10 effect in bone marrow-derived macrophages (BMDMs) in response to LPS stimulation. To grow bone-marrow derived macrophages (BMDMs), bone marrow cells were flushed from femur and tibia bones of IL-10-deficient mice strain in C57BL/6 background between 6 and 12 weeks of age, and were grown at 37°C in a humidified incubator in RPMI-1640 medium containing L-glutamine, 10% FCS and 30% L929 supernatant (i.e. BMDM growth media) for 7-8 days. The RNA sequencing analysis was performed separately in two groups of samples. Group 1: Il10-/- BMDMs were stimulated for 3 h without (control samples C1 and C2) or with 100 ng/ml of lipopolysaccharide (LPS)(samples C3 and C4), 100 ng/ml of IL-10 (samples D1 and D2) or both (samples D3 and D4). Group 2: Il10-/- BMDMs were stimulated without (control samples K1 and K2) or with 100 ng/ml of LPS (for 6 h: samples K3 and K4; for 12 h: samples K9 and K10), 100 ng/ml of IL-10 (for 6 h: samples K7 and K8; for 12 h: samples K13 and K14) or both (for 6 h: samples K5 and K6; for 12 h: samples K11 and K12).
Project description:Mammalian microRNAs (miRNAs) are emerging as key regulators of the development and function of the immune system. Here, we report a strong but transient induction of miR-155 in mouse bone marrow after injection of bacterial lipopolysaccharide (LPS) correlated with granulocyte/monocyte (GM) expansion. Demonstrating the sufficiency of miR-155 to drive GM expansion, enforced expression in mouse bone marrow cells caused GM proliferation in a manner reminiscent of LPS treatment. However, the mir-155-induced GM populations displayed pathological features characteristic of myeloid neoplasia. Extending possible relevance to human disease, miR-155 was overexpressed in the bone marrow of patients with acute myeloid leukemia (AML). Furthermore, miR-155 repressed a subset of genes implicated in hematopoietic development and disease. These data implicate miR-155 as a contributor to physiological GM expansion during inflammation and to certain pathological features associated with AML, emphasizing the importance of proper miR-155 regulation in developing myeloid cells during times of inflammatory stress. Keywords: genetic modification Overall design: Construct stable RAW264.7 mouse macrophage cell lines expressing mir-155 or empty vector. RNA is extracted and global gene expression analysis performed to identify mir-155 regulated mRNAs.
Project description:This SuperSeries is composed of the following subset Series: GSE19482: Transcriptional responses of human monocyte-derived macrophages (HMDM) to lipopolysaccharide (LPS) GSE19490: Transcriptional responses of mouse BMM and TEPM to lipopolysaccharide (LPS) GSE19765: Transcriptional responses of human monocyte-derived macrophages (HMDM) to lipopolysaccharide (LPS) - Illumina arrays GSE19766: Transcriptional responses of mouse bone marrow-derived macrophages (BMM) to lipopolysaccharide (LPS) - Illumina arrays Refer to individual Series
Project description:We have employed whole genome microarray expression profiling to identify differently expressed genes following stimulation of wild-type bone-marrow derived macrophages with Escherichia coli lipopolysaccharide (LPS). Macrophages were stimulated with LPS (100 ng/ml) for 6 hours and a signature was identified that distinguished between infected and control samples. Expression of several genes from this signature was quantified in the same RNA samples by real-time PCR, confirming the predicted macrophage response pattern. Overall design: Gene expression in macrophages was measured at 6 hours after stimulation Three independent experiments were performed.