Project description:Biologic functions involved in innate immune response of macrophages rely on the precise regulation of kinds of immune molecular. In the virus infection procession, the macrophages are activated following a tightly controlled genetic programme where specific sets of genes are up-regulated or down-regulated. We used microarrays to detail the global programme of gene expression underlying VSV infection and identified distinct classes of up-regulated and down-regulated genes during this process. Mouse peritoneal macrophages were selected with/without VSV infection for 8 hours for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain expression profiles. We selected macrophages according to VSV infection at two time-points: uninfected macrophage(control) and VSV infected for 8 hour macrophages(VSV).
Project description:Purpose: Characterize the gene expression profile of of peritoneal mouse macrophages in Endotoxic shock and Tolerance through RNA sequencing Methods: RNA sequencing of RNA from peritoneal macrophages in Endotoxic shock and Tolerance isolated by peritoneal lavage and FACS sorting (F4/80+ CD11b+) Results: Endotoxic shock and Tolerance peritoneal mouse macrophages display differential gene expression. Conclusions: Endotoxic shock and Tolerance peritoneal mouse macrophages display differential gene expression.
Project description:Colony Stimulating Factor 1(CSF1) is known to promote osteoclast progenitor survival but its role in regulating osteoclast differentiation and mature osteoclast function are less well understood. Macrophages have the potential to differentiate into osteoclasts and are also considered as osteoclast precursors. The microarray screen was designed to identify potential CSF1 targets in macrophage and osteoclast lineage. Wild type C57/BL6 mice were treated with 0.2 ml 10 % thioglycolate medium (FTG) / mouse by intra-peritoneal injection. Mice were sacrificed one week after injection followed by intra-peritoneal lavage with 15ml PBS. Macrophages were plated at a density of 1.5×10^6/well in 6-well plates and cultured in ?-MEM supplemented with 10% FBS. At 80% confluence, cells were treated with 100ng/ml CSF1 for 12 hours and RNA extracted. Microarray analysis was performed using Affymatrix Mouse Genome 430 2.0 Arrays. Data were analyzed by Gene spring 6.2 software.
Project description:To investigate the function of RFX1 in the regulation of macrophage polarization, we overexpressed RFX1 in mouse peritoneal macrophages(PMAs) by lentivirus infections. We then performed gene expression profiling analysis using data obtained from RNA-seq.
Project description:Setdb1 is one of the H3K9 methyltransferases and represses gene expression by H3K9 methylation. In an attempt to elucidate the role of Setdb1 in the TLR4-mediated inflammatory responses, we performed DNA microarray analysis using lipid A (the active component of LPS)-stimulated peritoneal macrophages from macrophage specific Setdb1 KO (KO) and WT mice. The genes upregulated by lipid A treatment in WT macrophages and further increased in KO macrophages contain many genes associated with interleukins and chemokines. Peritoneal macrophages from WT and KO mice were stimulated with lipid A 10 ng/ml or vehicle for 4 h. Microarray analysis was performed using Affymetrix Mouse 430 2.0.
Project description:Macrophage-inducible C-type lectin (Mincle, Clec4e) is a pathogen sensor that recognizes pathogenic fungi and Mycobactrium tuberculosis. We perfomed microarray analysis using peritoneal macrophages stimulated with TDM, a mycobacterial cell wall glycolipid that is known to be a Mincle ligand. Many chemokine and cytokine genes were upregulated in wildtype macrophages stimulated with TDM. Upregulation of these genes were completely abolishd in Mincle KO macrophages. Peritoneal macrophages from WT and Mincle KO mice were stimulated with TDM or vehicle for 24 h (3 samples each). Microarray analysis was performed using Affymetrix Mouse 430 2.0.
Project description:We have identified a number of miRNAs that are differentially expressed in LPS treated mouse peritoneal macrophages, as compared to untreated cells Peritoneal macrophages treated with LPS for 0, 4, 12, 24h. RNA was isolated and miRNA array assays were performed by Exiqon (miRCURY™ LNA Array version 10.0)