ABSTRACT: Hypoxia promotes macrophage polarization triggered by IL-6.In an attempt to gain speci?c insight into the mechanism of macrophage polarization during hypoxia. We used microarrays to detail the global programme of gene expression and identified distinct classes of changed genes during this process. Four groups of RAW264.7 cells were harvested for RNA extraction and hybridization on Affymetrix microarrays (normoxia control, normoxia+IL-6,hypoxia control,hypoxia+IL-6).
Project description:Hypoxia promotes macrophage polarization triggered by IL-6.In an attempt to gain specific insight into the mechanism of macrophage polarization during hypoxia. We used microarrays to detail the global programme of gene expression and identified distinct classes of changed genes during this process.
Project description:Purification of mouse mRNAs encoding signal transducer and activator of transcription 3 (Stat3) identified many miRNAs using multiplex miRNA array in RAW264.7 cells. To investigate the miRNAs targeting to mouse Stat3 gene in macrophage cells, we performed a protocol miRIP to affinity purify the miRNAs from an endogenous segment of Stat3 mRNA using nucleic acid hybridization and use multiplex miRNA array to identify the associated miRNAs.
Project description:Unanticipated regulatory protein modifications can be discovered from the unbiased comprehensive analysis of biological systems using SAMPEI. To explore this idea, we sought to map protein modifications induced during mammalian cell differentiation, modeled by the response of mouse RAW264.7 macrophage cells to lipopolysaccharide (LPS), a potent inducer of macrophage activation and differentiation that involves extensive protein and metabolic signaling.
Project description:Botulinum neurotoxin type A (BoNT/A) is one of the most potent protein toxins, which makes it a possible biological weapon and therapeutic. Using microarray analysis we performed global transcriptional profiling of RAW264.7 cells, a murine alveolar macrophage cell line. Time dependent expression profiles after treatment of 1nM or 5nM Botulinum neurotoxin A
Project description:Toxoplasma strains are known to inhibit the expression of several interferon-gamma induced genes, and a type II strain was shown to dysregulate genome-wide responses to interferon-gamma in human fibroblasts (Kim et al., 2007, J Immunol.). In this study we aimed to determine the effect of infection with three clonal lineages of Toxoplasma, type I, II, and III strains on genome-wide interferon-gamma induced transcription in murine macrophages. We also assessed the effect of the two main Toxoplasma modulators of mouse macrophage transcription, ROP16 and GRA15 (Jensen et al., 2011, Cell Host Microbe). We used Affymetrix microarrays to analyze host cell transcription after Toxoplasma infection and interferon-gamma stimulation. RAW264.7 murine macrophages were left uninfected or infected with type I (RH), type I ?rop16 (RH ?rop16), type II (Pru), type II ?gra15 (Pru ?gra15), or type II (CEP) parasites at an MOI ~5 for 18 hours and subsequently stimulated with murine IFN-? for six hours. Plaque assays were done to assess parasite viability. Total RNA was isolated and hybridized to Affymetrix Mouse 430A 2.0 gene chips.
Project description:RAW264.7 cell was stably transfected with Tim-3 siRNA(SI) or negative control(NC) and were analyzed for microRNA expression Differerently expressed microRNA in Tim-3 knockdown RAW264.7 cells were summarized and were used for further analysis RAW264.7 cells stably transfected with Tim-3 siRNA(SI) or negative control(NC) and were used to collect microRNA without any other treatment
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to evaluate the gene expression of M2-polarized RAW264.7 cells compared with M0 RAW264.7 cells. Methods: RAW264.7 cells were polarized toward M2 using IL-4. M0 RAW264.7 cells were maintained in culture without IL-4. Total RNA of M2 and M0 RAW264.7 cells was extracted. RAW264.7 cells RNA profiles were generated by deep sequencing for two groups (M2 versus M0 RAW264.7 cells) with three samples each. Results: There were significant differences between M2 and M0 RAW264.7 cells. Conclusions: Polarization of RAW264.7 cells from M0 to M2 induces various changes at the transcription level.
Project description:Investigation of whole genome gene expression level changes in a Tim-3 knockdown RAW264.7 cell, compared to control siRNA transfected RAW264.7 cell. The Tim-3 knockdown of this cell render it hyper-reactive to TLR stimulation. Tim-3 knockdown analyzed in this study are further described in Li Y, Feng J, Geng S, Geng S, Wei H, Chen G, Li X, Wang L, Wang R, Peng H, Han G, Shen B, Li Y. 2011. The N- and C-terminal carbohydrate recognition domains of galectin-9 contribute differently to its multiple functions in innate immunity and adaptive immunity. Mol Immunol. 48(4):670-7. (PID 21146220) A six chip study using total RNA recovered from Tim-3 knockdown and control RAW264.7 cells. There are 135,000 probes in a chip,each chip measures the expression level of 27526 genes of mouse from Nimblegen. Fold-change screening between the two groups obtained from the experiment. The threshold used to screen up or down regulated genes is Fold changeM-oM-<M-^-oM-<M-^]2.0
Project description:The purpose of this study was to determine what are the effects of Src deficiency on innate antiviral response upon virus infection in RAW264.7 cells. Wild type and Src-/- RAW264.7 cells were infected with vesicular stomatitis virus (VSV) or herpes simplex virus 1 (HSV-1) for 6h. Then the differentially regulated genes were analyzed. Wild type and Src-/- RAW264.7 cells were infected with vesicular stomatitis virus (VSV, MOI=1) or herpes simplex virus 1 (HSV-1, MOI=5) for 6h. Equal amounts of RNA were assayed for gene expression using Affymetrix mouse 430 2.0 arrays.