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: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: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: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:Endothelial cells derived from freshly purified human islets were incubated with microvesicles collected by ultracentrifugation of supernatants of endothelial progenitor cells (EPCs) isolated from peripheral blood of healthy volunteers.
Project description:Male Sprague Dawley adult rats were subjected to a cervical hemisection at C2 (C2HS). Costal diaphragm (both ipsilesional and contralesional sides) were assessed under control conditions (sham surgery) and at 1 and 7 days post-C2HS. We used SA Biosciences Rat Skeletal Muscle Development and Disease RT2 Profiler PCR Array to quantitate gene expression of muscle atrophy and regeneration-relevant genes from the uninjured and injured tissues on the indicated sides and at the indicated timepoints post-lesion. qPCR gene expression profiling. Tissue was derived from the costal diaphragm from control animals, and from contralesional and ipsilesional sides at days 1 and 7 post-lesion. RNA was extracted from flash-frozen tissue (TRIzol reagent) and quantitated via spectrophotometry. Equal amount total RNA from each donor was pooled prior to gene expression analysis. N=3 or 4 rats per condition. Four genes in the array study appeared to undergo unphysiologic increases in expression following SCI: Mmp9, Leptin, activin A and alpha actinin. For these genes, expression levels in control tissue were detectable but extremely low, thus fold-increases following SCI are numerically exaggerated. Therefore, we cannot explicitly comment on the physiologic scale of this response
Project description:Wild type (WT) and Pglyrp1-/- mice were treated with PBS or sensitized 5 days/week for 3 or 5 weeks with 10 µl per application of 2.5 mg/ml of purified house dust mite allergen. 3 days after the last sensitization the lungs were removed and homogenized, and RNA was isolated from the right lobes using the TRIZOL method. Quantitative reverse transcription real-time PCR (qRT-PCR) was used to quantify the amounts of mRNA in the lungs using custom RT2 Profiler PCR Arrays designed by us and manufactured by Qiagen/SA Biosciences. qRT-PCR gene expression profiling
Project description:MDA-MB-231 breast cancer cells were infected with either Ad-GFP, Ad-FLI1, or Ad-PDEF qPCR gene expression profiling of MDA-MB-231 breast cancer cells were infected with either Ad-GFP, Ad-FLI1, or Ad-PDEF.
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