Project description:Microglia are brain-resident, myeloid cells that play important roles in health and brain pathologies. While data on chromatin accessibility in activated murine microglia are available, there is no such data on rat microglia subjected to different stimuli. Herein, we report a comprehensive, replicated, FDR-controlled dataset of DNase-hypersensitive (DHS) open chromatin regions for the rat microglia. We compared open chromatin landscapes in untreated primary microglial cultures and cultures stimulated for 6h with either lipopolysaccharide (LPS) or glioma-conditioned medium (GCM). Glioma-secreted factors induce the pro-invasive and immunosuppressive activation of microglia, in which these cells promote tumor growth. The open chromatin landscape of the rat microglia consisted of 126,640 reproducible DHS regions, of which 12,357 and 2,303 significantly changed openness following the stimulation with LPS and GCM, respectively. Active genes had constitutively open promoters, but there was no direct dependence between cumulative openness of DHS regions near to a gene and its expression. LPS-regulated DHS regions were more frequent in introns, while GCM-regulated regions were more frequent away from gene bodies. GCM and LPS treatment differentially affected open chromatin regions mapped to genes in the pathways of Toll-like-receptor signaling and the Axon guidance, suggesting that the molecular machinery used by migrating microglia is similar to that of growing axons and, moreover, that modulation of these pathways is instrumental for glioma to induce a pro-invasive polarization of microglia. Our dataset of open chromatin regions active in rat microglia will constitute a useful resource for studies of the transcriptional regulation in this system.
Project description:Several independent microglia preparations performed on different days were plated onto wells of 24-well plates, and either left untreated (control), treated with the C6 glioma-conditioned medium, or treated with LPS.<br>For each culture condition, a number of RNA samples were prepared, with each sample then separately labeled and hybridized. These RNA samples were prepared from non-overlapping sets of wells from at least two microglia preparations, so they constitute biological replicates.
Project description:Several independent microglia preparations performed on different days were plated onto wells of 24-well plates, and either left untreated (control), treated with the C6 glioma-conditioned medium (GCM). RNA samples were prepared at 6, 24 and 48 hours from the start of the treatment, as well as from the control untreated microglia, with each sample then separately labeled and hybridized. These RNA samples were prepared from from at three independent microglia preparations, so they constitute biological replicates.
Project description:To study the effect of myoblast-derived paracrine factors on cardiomyocyte gene expression, microarray analysis was performed from isolated rat fetal cardiomyocyte cultures. These cultures were treated for 24 hours with fresh culture medium (control), skeletal myoblast-conditioned medium, or cardiac fibroblast-conditioned medium.
Project description:The protein(s) larger than 100 kDa in conditioned medium from lung cancer cell lines activated microglia. The >100 kDa fraction of conditioned medium from H460, H1299, H2030, or H292 cells was collected. Proteins present in the conditioned medium were analyzed by LC-MS/MS.
Project description:Primary human skeletal muscle cells (Lonza) were treated with LLC1 conditioned medium, LLC1 conditioned medium plus Calcitriol, LLC1 non-conditioned medium or LLC1 non-conditioned medium plus Calcitriol for a period of 24 hours prior to isolation of RNA.
Project description:To study the effect of myoblast-derived paracrine factors on cardiomyocyte gene expression, microarray analysis was performed from isolated rat fetal cardiomyocyte cultures. These cultures were treated for 24 hours with fresh culture medium (control), skeletal myoblast-conditioned medium, or cardiac fibroblast-conditioned medium. In total, 3 samples were analyzed.
Project description:The aim of the experiment was to identify differences in gene expression in the microglia due to the presence of a brain tumor, at 14 days after a stereotactic injection of 5 x 10E4 rat C6 glioma cells into the right striatum of the rat. The control (naive) animals were left untreated. The brain hemispheres from the control animals, or the tumor-bearing hemispheres were dissected, gently dissociated into cells and the microglia (CD11b+CD45low cells) were sorted out.