Project description:Tubular endothelial cells were cultured in hypoxic conditions (1% O2) and treated or not with microvesicles derived from endothelial progenitor cells.<br>mRNA profiling of hypoxic cells, treated or not with MVs, was analyzed after nromalization with mRNA profiling of normoxic cells.<br><br>This experiment was updated on 27th Jan 2011 to correct descriptions.
Project description:Identification of transcriptional profile of several genes involved in diabetes in islet-derived extracellular vesicles (Evs). Recently, EVs are identified as a new mechanism in cell-to-cell communication by transfer of protein and genic information (mRNA, microRNA). Their role is under investigation in immunology, stem cell and cancer, but not in islets and diabetes. The aim of this experiment is to identify mRNA transcripts (in particular, mRNA transcripts involved in diabetes pathophysiology) present in islet Evs.
Project description:HUVECs were cultured in hypoxic conditions for 24 hours and treated or not with 1uM caffeic acid. Then cells were evaluated for expression of different genes involved in angiogenesis
Project description:Glomerular endothelial cells were cultured in normal condition and treated or not with microvesicles derived from endothelial progenitor cells. mRNA profiling of glomerular endothelial cells , treated with MVs, was analyzed after normalization with mRNA profiling of untreated cells.
Project description:TLR3, TLR7, and TLR9 stimulation induces many mouse inflammatory and autoimmune cytokines or immune receptors DRGN were cultures 5 days prior to a 16 hour stimulation - Three separate studies were analyzed for inflammatory response
Project description:Heterogeneity of lung tumor endothelial cell (TEC) phenotypes across patients, species (human/mouse) and models (in vivo/vitro) remains poorly inventoried at the single-cell-level. We single-cell RNA-sequenced 56,771 ECs from human/mouse (peri)-tumoral lung and cultured human lung TECs, detected 17 known and discovered 16 novel phenotypes, including TECs presumably regulating immune surveillance. We resolved the canonical tip TECs into a known migratory tip and a novel basement-membrane remodeling breach phenotype. Tip-TEC signatures correlated with patient-survival, and tip/breach TECs were most sensitive to VEGF-blockade. By similarity analysis, only tip-TECs were congruent across species/models and shared conserved markers. Integrated analysis of the scRNA-seq data with orthogonal multi-omics and meta-analysis data across different human tumors, validated by functional analysis, identified collagen-modification as angiogenic candidate pathway.
Project description:Murine macrophages were isolated from the lungs of mice given a pulmonary challenge with C. neoformans strain H99. Mice were either given a protective (H99γ) or a mock (HKCn) immunization prior to C. neoformans H99 challenge, and macrophages were isolated from the lungs of mice 24 hours, 3 days, or 7 days post-challenge using anti-CD11b microbeads according to the Miltenyi cell sorting system. We used SA Biosciences Toll-like Receptor PCR assay panel to quantitate gene expression of signal transduction factors in total RNA isolated from macrophages derived from immunized mice compared to non-immunized. qPCR gene expression profiling. Macrophages from 5 mice per group were pooled and assayed as indicated in the summary. Each experiment was performed 3 times and the resulting Ct values of each group from each experiment averaged prior to data analysis. TIme points were analyzed separately
Project description:Numerous studies have suggested a link between fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) signaling pathways; however the nature of this link has not been established. To evaluate this relationship we investigated VEGF signaling in endothelial cells with disrupted FGF signaling in vitro and in vivo. We find that endothelial cells lacking FGF signaling become unresponsive to VEGF due to down regulation of VEGFR2 expression caused by reduced Vegfr2 enhancer activation, which is in turn caused by reduced activation of Ets family transcription factors. In vivo this manifests in the loss of vascular integrity and morphogenesis. Thus, basal FGF stimulation of the endothelium is required for maintenance of VEGFR2 expression and the ability to respond to VEGF stimulation and accounts for the hierarchic control of vascular formation by FGFs and VEGF. Primary mouse lung endothelial cells were transduced with either Adeno-Null (empty) or Adeno- dominant negative FGF receptor 1 and harvested 24 hours after transduction. Total RNA was extracted and subjected to the analysis using SuperArray GEArray Q Series Mouse Angiogenesis Gene Array. Comparisons were made between treatments.
Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A) from SABiosciences. Mice used in this study were 2 week old males. Total RNA was extracted from whole cerebella of 2 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A).