Fatostatin/Saracatinib drug treatment of mouse primary microglia cultures
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PROVIDER: PRJNA612200 | ENA |
REPOSITORIES: ENA
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Project description:Saracatinib drug treatment of mouse primary microglia cultures
| PRJNA612216 | ENA
Project description:To test the effect of saracatinib on gene expression patterns in microglia, we treated mouse (C57BL/6) primary microglial cultures with saracatinib (1microM, 24h) versus DMSO vehicle control and quantified gene expression. Sequencing libraries were prepared using the Lexogen QuantSeq 3'Forward kit with sequencing performed on an Illumina HiSeq 4000 (single end reads 1 x 65; 5M reads per sample). For processed data, reads were aligned against GRCm38 and quantified using STAR. Samples were normalized with CQN. Batch correction was completed using ComBat from sva version 3.6.0.
2020-11-05 | GSE146865 | GEO
Project description:Fatostatin drug treatment of mouse primary microglia cultures
| PRJNA612217 | ENA
Project description:To test the effect of fatostatin on gene expression patterns in microglia, we treated mouse (C57BL/6) primary microglial cultures with fatostatin (5microM, 72h) versus DMSO-only control and quantified gene expression using 3' end labeling. Sequencing libraries were prepared using the Lexogen QuantSeq 3'Forward kit and sequencing of single end reads (1 x 65) was performed on an Illumina HiSeq 4000 at 5M reads per sample. For processed data, reads were aligned against GRCm38 using STAR. Transcripts were quantified and annotated against GencodeM11 using Rsubread. Samples were normalized with CQN.
2020-11-05 | GSE146864 | GEO
Project description:Preparation of primary microglial cultures from postnatal mice is tedious with a low yield, high variability and risk of astrocytic contamination. Microglia derived from embryonic stem cells (ESdM) have been suggested as alternative source, but it is unclear how closely ESdM resemble the molecular phenotype of primary microglia. Here, we performed a whole transcriptome analysis of ESdM in comparison to primary cultured and flow cytometry-sorted microglia and compared the microglial transcriptome to other cell types. Cultured microglia and ESdM were related to sorted microglia, but clearly distinct from other myeloid cell types, T cells, astrocytes and neurons. ESdM and primary cultured microglia showed strong overlap in their transcriptome. Only 144 gene transcripts were differentially expressed between both cell types, mainly derived from immune-related genes with a higher activation status of pro-inflammatory and immune defense genes in primary microglia compared to ESdM. Flow cytometry analysis of cell surface markers CD54, CD74 and CD274 selected from the microarray confirmed the close phenotypic relation between ESdM and primary cultured microglia. Thus, assessment of genome-wide transcriptional regulation demonstrates that microglia are distinct from other macrophage cell types and that mouse pluripotent stem cell-derived microglia are closely related to cultured postnatal microglia. Comparison of different primary neuronal cells with ES-cell derived microglial cells
2013-07-11 | E-GEOD-43808 | biostudies-arrayexpress
Project description:phosphopeptides were isolated from primary microglia, and identified by LC-MS/MS analysis.
2023-02-24 | PXD040217 | JPOST Repository
Project description:This SuperSeries is composed of the SubSeries listed below.
2020-11-05 | GSE146866 | GEO
Project description:Effect of GW2580 treatment on primary microglia transcriptome
| PRJNA769680 | ENA
Project description:Preparation of primary microglial cultures from postnatal mice is tedious with a low yield, high variability and risk of astrocytic contamination. Microglia derived from embryonic stem cells (ESdM) have been suggested as alternative source, but it is unclear how closely ESdM resemble the molecular phenotype of primary microglia. Here, we performed a whole transcriptome analysis of ESdM in comparison to primary cultured and flow cytometry-sorted microglia and compared the microglial transcriptome to other cell types. Cultured microglia and ESdM were related to sorted microglia, but clearly distinct from other myeloid cell types, T cells, astrocytes and neurons. ESdM and primary cultured microglia showed strong overlap in their transcriptome. Only 144 gene transcripts were differentially expressed between both cell types, mainly derived from immune-related genes with a higher activation status of pro-inflammatory and immune defense genes in primary microglia compared to ESdM. Flow cytometry analysis of cell surface markers CD54, CD74 and CD274 selected from the microarray confirmed the close phenotypic relation between ESdM and primary cultured microglia. Thus, assessment of genome-wide transcriptional regulation demonstrates that microglia are distinct from other macrophage cell types and that mouse pluripotent stem cell-derived microglia are closely related to cultured postnatal microglia.
2013-07-11 | GSE43808 | GEO
Project description:Microglia play important and dynamic roles in mediating a variety of physiological and pathological processes during the development, normal function, and degeneration of the central nervous system. We report here the development of a SILAC-labeled model system suitable for mass spectrometry-based proteomic profiling in primary rat microglia. A mixed primary glia culture was first established by isolating newborn brain cells from Sprague-Dawley rats and maintaining the culture in a fetal bovine serum-supplemented and chemically defined media containing either the regular amino acids (light label media) or stable isotope-labeled amino acids (heavy label media). After isolation of microglia from the mixed glia cultures, mass spectrometric analysis indicated that the incorporation efficiency of the heavy-labeled amino acids was between 70-80%. Upon stimulation with bacterial endotoxin lipopolysaccharide to induce classical (M1) activation, the light- and heavy-labeled primary microglia responded indistinguishably as judged by the up-regulation of inducible nitric oxide synthase. More important, the relative quantitation accuracy and proteome coverage obtained by the reported SILAC model allowed for identification of classical microglial activation markers and pathways involved in the proinflammatory response of microglia as determined by bioinformatic analysis. The establishment of this viable primary microglia SILAC model will further expand our capacity for global scale proteomic profiling of pathways mediating the multifaceted microglial responses in various physiological and pathological processes.
2016-04-07 | PXD002759 | Pride