Project description:In order to identify the genes regulated in mouse embryonic stem cells (mESC) by the effect of low concentrations of nitric oxide (NO), we analysed the transcriptome of cells treated with NO and compared it to those of cells cultured in the absence of leukemia inhibitory factor (LIF), and in the presence of LIF. We used the cell line D3-pOct4, which carries the enhanced Green Fluorescence Protein gene (eGFP) under the control of the Oct-4 promotor. This line is continuously maintained in the undifferentiated state in the presence of LIF, in comparison with the wild type line .
Project description:In order to identify the genes regulated in mouse embryonic stem cells by the effect of low concentrations of nitric oxide (NO), we analysed the transcriptome of cells treated with NO and compared it to those of cells cultured in the absence of leukemia inhibitory factor (LIF), and in the presence of LIF. We used the cell line D3-pOct4, which carries the enhanced Green Fluorescence Protein gene (eGFP) under the control of the Oct-4 promotor. This line is continuously maintained in the undifferentiated state in comparison with the wild type line in the presence of LIF. Experiment Overall Design: D3-pOct4 cells cultured in the presence of 1000 U/ml of LIF, in the absence of LIF, and in the absence of LIF with 2uM of diethylenetriamine nitric oxide adduct (DETA)/NO were grown for 7 days, maintaining the conditions of the treatment everyday. DETA-NO (Sigma) was used as donor of nitric oxide. RNA was extracted and analyzed by Bioanalyzer of Agilent. The cDNA microarray chip used was Mouse Genome 430_2.0 (Affymetrix) and the data were analyzed with GeneChip Operating System v1.4.0.036 using global scaling as the normalization method.
Project description:Identification of genes regulated in human embryonic stem cells by the effect of low concentrations of nitric oxide. The transcriptome of cells treated with NO were compared with that of cells cultured in abscense of LIF, and undifferentiated ES cells cultured in presence of LIF. The H181 hES cell line was used for the experiments. These data complement a previous study realized in mES cells under the same NO conditions . The aim of the work is to know how NO modify transcriptome of ES cells and how participate in the modulation of self renewal and differentiation.
Project description:Identification of genes regulated in human embryonic stem cells by the effect of low concentrations of nitric oxide. The transcriptome of cells treated with NO were compared with that of cells cultured in abscense of LIF, and undifferentiated ES cells cultured in presence of LIF. The H181 hES cell line was used for the experiments. These data complement a previous study realized in mES cells under the same NO conditions . The aim of the work is to know how NO modify transcriptome of ES cells and how participate in the modulation of self renewal and differentiation. Passages between 72 and 74 of the H181 cell line were cultured on Matrigel-coated tissue culture plates in presence of 8ng/ml of bFGF for maintain them undifferentiated (control). For treatments, cells were cultured in the absence of bFGF to induce differentiation and treat cells with 2uM of DETA/NO. Cells were grown for 6 days mantaining the conditions of the treatment each 19 hours. Dietilentriamine-NO (DETA-NO) (Sigma) was used as donor of nitric oxide. Expression data of cells treated with NO in absence of bFGF were compared with cells cultured in absence of this, in order to observe how the treatment affects the transcriptome, and if there is some similarity with undifferentiated condition in presence of bFGF.
Project description:While in transit within and between hosts, uropathogenic E. coli (UPEC) encounter multiple stresses, including substantial levels of nitric oxide and reactive nitrogen intermediates. Strains of UPEC become conditioned to high concentrations of acidified sodium nitrite (ASN), a model system used to generate nitrosative stress. We used microarrays to define the expression profile of UPEC that have been conditioned for growth in ASN.
Project description:The goal of this study was to measure the effects of nitric oxide exposure (using DETA NONOate as a nitric oxide donor) on transcription in Caulobacter. Untreated Caulobacter crescentus were grown to a density of 0.3 (at OD660) in PYE medium (pH 7) in rolled culture tubes. DETA-NONOate treated Caulobacter crescentus were grown to a density of 0.3 (at OD660), and then treated with 100 mM DETA NONOate for 30 minutes.
Project description:Asymmetric dimethylarginine (ADMA) is a naturally occurring inhibitor of nitric oxide synthesis that accumulates in wide range of diseases associated with endothelial dysfunction and enhanced atherosclerosis. Plasma ADMA has been implicated as a major novel cardiovascular risk factor, but the mechanisms by which low concentrations of ADMA produce adverse effects on the cardiovascular system are unclear. We have treated human coronary artery endothelial cells with ADMA at 2uM (a pathophysiological dose) and 100uM (a pharmacological dose), for 24h.
Project description:Background: We would like to use Drosophila cell lines to investigate the transcriptional responses to Nitric Oxide donors and inhibitors. As a pilot to this we would like to compare the transcriptional profile of 3 cell lines we are using in our lab (S3, Kc and S3). This study is being done in parallel to an in vivo genetic screen to indentify required genes. This work is funded by the BBSRC. Plan: Pairwise comparisons of each cell line to the other two. Cells will be grown at low density in identical conditions. Cells will be harvested and RNA prepared in Trizol as recommended.
