Transcription profiling of mouse naive T cells compared to regulatory T cells and activated T-cells induced to express Foxp3
ABSTRACT: CD4+CD25-CD62L+ naive T-cells were FACS isolated from wild type mouse lymph node. These were compared to regulatory T cells (Treg; CD4+C25+) and activated T-cells induced to express Foxp3 by treatment with rapamycin and LY294002 (Treg-like cells). RNA was extracted from duplicate cultures using RNA-Bee, labelled with the Affymetrix GeneChip Small Sample protocol and hybridised to Affymetrix Mouse 430 2.0 Arrays.
Proceedings of the National Academy of Sciences of the United States of America 20080528 22
Regulatory T (Treg) cells safeguard against autoimmunity and immune pathology. Because determinants of the Treg cell fate are not completely understood, we have delineated signaling events that control the de novo expression of Foxp3 in naive peripheral CD4 T cells and in thymocytes. We report that premature termination of TCR signaling and inibition of phosphatidyl inositol 3-kinase (PI3K) p110alpha, p110delta, protein kinase B (Akt), or mammalian target of rapamycin (mTOR) conferred Foxp3 expr ...[more]
Project description:Mononuclear cells were isolated from granulocyte colony stimulating factor mobilised peripheral blood. CD34+ cells were selected and separated into adherent and non-adherent cells. Adherent and non-adherent samples were amplified and labelled using two IVT cycles, and gene expression profiling was performed by hybridisation to Affymetrix Human Genome U133 Plus2.0 Arrays.
Project description:Title: Regulation of gene expression through mitogen-activated protein kinase cascades in cardiac myocytes.<br/> Description: The aim of this study is to identify the changes in gene expression induced in rat neonatal <br/> ventricular myocytes, a well-established cell culture model, by endothelin-1, <br/> a known hypertrophic agonist, and to determine which of the changes are <br/> mediated through the ERK cascade. This continues TKACs previous study of the <br/> effects of oxidative stress, which induces cardiac myocyte apoptosis.<br/>
Project description:Title: Regulation of gene expression through mitogen-activated protein kinase cascades in cardiac myocytes.<br/> Description: We aim to identify the changes in gene expression induced in rat <br/> neonatal ventricular myocytes, a well established cell culture model, <br/> by phenylepherine, a known hypertrophic agonist, and to determine which <br/> of the changes are mediated through the ERK cascade. In addition, we <br/> intend to extend the previous studies (EXP_TKAC_0102_01 and <br/> EXP_TKAC_1103_02) on endothelin-1 induced hypertrophy and the oxidative <br/> stress induced by H2O2 by using additional concentrations and timepoints.
Project description:Title: Gene Expression analysis of the role of RIP140 in adipocyte differentiation<br/> Description: RIPKO cells are mouse embryo fibroblasts (MEF) that were derived from <br/> RIP140-knockout mice and have been established as a RIP140 null cell line,<br/> while RIPKO-L7 and RIPKO-L16 are RIPKO cells for which the expression of<br/> RIP140 has been reintroduced using a lentiviral expression system. <br/> Using an established standard cocktail of hormones that includes IBMX, <br/> insulin, dexamethasone and a PPAR gamma agonist the cells were <br/> differentiated into adipocytes and RNA isolated at day 0 and day 10 <br/> after the addition of the cocktail.
Project description:Title: Comparison of ovarian gene expression in RIP140 wild type, heterozygous and knock out mice.<br/> Description: RIP140 null mice fail to ovulate. This experiment was designed to <br/> determine the gene expression profile of these animals compared <br/> to wild type and heterozygous animals following hormone treatments <br/> that induce the biological changes required for ovulation to occur.
Project description:Ventricles were removed from 2-day Sprague_Dawley rat litters from which primary cardiac myocytes (>95%) were isolated. Myocytes were either untreated or exposed to either endothelin-1 for 30, 60 or 120 minutes, cycloheximide for 70 or 130 minutes, endothelin-1 for 10 minutes followed by cycloheximide for 70 minutes, azekenpaullone for 60 minutes before extraction of total or polysomal (actively transcribed) RNA. Labelled cRNA was hybridised to Affymetrix Rat 230 2.0 arrays.
Project description:Undifferentiated mouse embryonic stem cells stably expressing the HOXB1 transcription factor under doxycycline tet-on control were differentiated into embryoid bodies. Nestin+ neuroepithelial cells were selected by transfer of the embryoid bodies to ISTFn media. Expression of HOXB1 was induced by addition of doxycycline or not induced on the seventh day of selection and also for one day after trypsination and transfer to laminin/polyornithine substrate on matrigel substrate.
Project description:Title: Gene expression profiling of human heart samples: a comparison between normal and hibernating myocardium.<br/> Description: The aim of this project is to investigate the changes in global gene<br/> expression in hibernating,(preserved wall thickness, impaired <br/> contractility at rest, recruitable with low dose dobutamine <br/> and 50% transmural hyperenhancement with gadolinium) and normal myocardium,<br/> (preserved myocardial wall thickness, normal contractility, no significant improvement in functional recruitability with dobutamine and no cardiac fibrosis seen).
Project description:Title: Gene Expression analysis of naive CD8+ T-cell differentiation during a primary immune response Description: C57BL/6 wild type mice are administered GK1.5 (depleting anti CD4 antibody) or isotope control by intraperitoneal injection at 3 and 1 day pre-infection, then every 36-48 hour post infection. 24 hours pre-infection, splenocytes from F5 RAG -/- mice are stained with CFSE and 10^7 splenocytes are transfered to recipient C57BL/6 mice by intravenous injection. 24 hours laters, recipient mice are infected with 10^7 PFU of a recombinant Vaccinia virus expressing NP366-74. At timepoints 0, 12, 24, 48, 72 and 96 splenocytes are obtained and FACS sorted to obtain CD8+ T-cells.
Project description:A mouse leydig tumour cell line (mLTC-1) was grown in culture and stimulated with either human chorionic gonadotropin (hCG, 10 ng/ml) alone or hCG & bisphenol A (BPA, an endrocrine disrupting chemical, 10 uM) for three hours. Cells from triplicate plates were harvested, RNA extracted, labelled and hybridised to Affymetrix Mouse genome 430 2.0 arrays. Preliminary work by RT-PCR had shown that stimulation of the cell line by BPA resulted in changes in gene expression in the spermatogenesis pathway at this time point.