Project description:We have identified a number of miRNAs that are differentially expressed in LPS and IFNγ treated RAW264.7 cells, as compared to untreated cells. Microarray and quantitative real-time PCR (qRT-PCR) results showed that miR-103 decreased while the STAT1 expression increased substantially in RAW264.7 derived M1 macrophage, and there was a significant negative correlation between miR-103 and STAT1.Overexpression of miR-103 suppressed M1 polarization by inhibiting STAT1/IRF1 signaling pathway and vice versa in vitro. STAT1 is a direct target of of miR-103.
Project description:The goal of this experiment was to validate a classification model built on other bulk transcriptomic data that was capable of identifying in vitro stimulated M-CSF differentiated macrophages. Specifically, the classification model was capable of distinguishing: M-LPSearly (2-4 hours) M-LPSlate (18-24 hours) M-LPS+IFNγ M-IFNγ M-IL4 M-IL-10 M-dex
Project description:We sought to characterize the genomic localization of the YEATS domain protein AF9 in the macrophage-like cell line RAW 264.7 +/- LPS stimulation and +/- sodium crotonate pretreatment. We performed chromatin immunoprecipitation followed by massively parallel sequencing (ChIP-seq) for endogenous AF9 or for a FLAG-tagged AF9 transgene in RAW264.7 or a RAW264.7 cell line engineered to express FLAG-AF9, respectively.
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 M1-polarized RAW264.7 cells compared with M0 RAW264.7 cells. Methods: RAW264.7 cells were polarized toward M1 using LPS and IFN-γ. M0 RAW264.7 cells were maintained in culture without LPS and IFN-γ. Total RNA of M1 and M0 RAW264.7 cells was extracted. RAW264.7 cells RNA profiles were generated by deep sequencing for two groups (M1 versus M0 RAW264.7 cells) with three samples each. Results: There were significant differences between M1 and M0 RAW264.7 cells. Conclusions: Polarization of RAW264.7 cells from M0 to M1 induces various changes at the transcription level.
Project description:Nuclear interaction studies by ChIP coupled with mass spectrometry identified the COMPASS/SETD1A complex as interaction partner of the glucocorticoid receptor (GR) in murine bone marrow-derived macrophages (BMDMs). Here, we profiled H3K4me1, H3K4me2 and H3K4me3 in wild-type and Setd1a hypermorphic (Setd1aDel/+) Raw264.7 cells after LPS and Dex+LPS stimulation by spike-in ChIP-Seq.
Project description:After stimulation of RAW264.7 cells with LPS and Harmine, high-throughput sequencing technology was used to explore the changes in the inflammation level of RAW cells and the enrichment of signaling pathways after LPS and Harmine treatment, and to find an interesting target pathway.
Project description:RAW264.7 cells were transfected with Synbindin siRNA for 48hours, then 100ng/ml LPS were treated cells for 4 hours. Before and after LPS stimulation, cells were collected and RNA were extracted for RNA-seq.
Project description:We report the miRNA profiles of macrophages (cell line: RAW264.7) and endothelial cells (cell line: TIME) after supplementation with polyunsaturated fatty acids (PUFA) and stimulation with LPS/pro-inflammatory cytokines. The fatty acids docosahexaenoic acid (DHA, C22:6n3) or arachidonic acid (AA, C20:4n6) were included in the culture medium in concentrations of 15 µmol/L using ethanol as a vehicle (0.2 % v/v final ethanol concentration). Cells were cultured in the enriched media totaling either 72 h (RAW264.7) or 144 h (TIME). Stimulation of cells was performed in the last 24 h of fatty acid supplementation by addition of either LPS (1 µg/mL; from E. coli serotype 0111:B4) for cell line RAW264.7 or the cytokines IL-1β, TNF-α, and IFN-γ each in a concentration of 5 ng/ml for cell line TIME.