Project description:Expression data from NIH-3T3 cells treated with mock, 100 U/ml IFN alpha or 100 U/ml gamma for 1 or 3h on nt-RNA labeled for 30-60 min at different times of interferon treatment Differential gene expression caused by IFN alpha or gamma was analyzed in newly transcribed RNA (nt-RNA) of NIH-3T3 cells treated for 1 or 3 h. RNA was labeled for 30 to 60 min and separated from total cellular RNA (tc-RNA) following Trizol RNA preparation and thiol-specific biotinylation We used microarrays to analyze the effects of IFNalpha and gamma treatment in newly transcribed RNA (nt-RNA) Keywords: time course
Project description:This SuperSeries is composed of the following subset Series:; GSE9973: Half-life determination for human B-cells (BL41); GSE9975: newly transcribed RNA (nt-RNA) for IFN alpha and gamma time course; GSE9977: Expression data from NIH-3T3 cells treated with mock, 100 U/ml IFN alpha or 100 U/ml gamma for 1or 3h; GSE10011: Expression data from NIH-3T3 cells used for half-life determination Experiment Overall Design: Refer to individual Series
Project description:Expression data from NIH-3T3 cells treated with mock, 100 U/ml IFN alpha or 100 U/ml gamma for 1 or 3h on nt-RNA labeled for 30-60 min at different times of interferon treatment; Differential gene expression caused by IFN alpha or gamma was analyzed in newly transcribed RNA (nt-RNA) of NIH-3T3 cells treated for 1 or 3 h. RNA was labeled for 30 to 60 min and separated from total cellular RNA (tc-RNA) following Trizol RNA preparation and thiol-specific biotinylation; We used microarrays to analyze the effects of IFNalpha and gamma treatment in newly transcribed RNA (nt-RNA) Experiment Overall Design: NIH-3T3 cells (5th to 15th passage after thawing) were split 24 h before start of the experiment. When starting the experiment 80% confluency was reached. The experiment was started by applying fresh, prewarmed, CO2-equilibrated medium containing either mock, 100U/ml IFN alpha or 100 U/ml IFN gamma. Labeling was either started simultaneously, 30 or 150 minutes later.
Project description:Differential gene expression caused by 1h and 3h of IFN alpha or gamma treatment was analyzed in total cellular RNA of NIH-3T3 cells compared to mock We used microarrays to analyze the effects of 1 and 3h of IFNalpha and gamma treatment in total cellular RNA Keywords: time course in total cellular RNA (tc-RNA)
Project description:G-protein coupled receptors (GPCRs) have diverse roles in physiological processes, including immunity. Gs-coupled GPCRs increase while Gi-coupled ones decrease intracellular cAMP. Previous studies suggest that, in epithelial cells, Gs-coupled GPCRs enhance whereas Gi-coupled GPCRs suppress pro-inflammatory immune responses. In order to examine the issue, we chose beta2 adrenergic receptor and GPR40 as representatives of Gs- and Gi- coupled GPCRs, respectively, and examined their effects on TNF-alpha and IFN-gamma-(TNF-alpha + IFN-gamma) induced gene expression by HaCaT. We used microarrays to detail the global changes of gene expression induced by a beta2 adrenergic receptor agonist terbutaline or GPR40 agonist GW9508 pre-treatment in TNF-alpha + IFN-gamma - stimulated HaCaT cells. HaCaT cells were pre-treated with terbutaline or GW9508, TNF-alpha + IFN-gamma were then added, and cultured for another 24 h. Cells were then used for RNA extraction and hybridization on Affymetrix microarrays. We sought to clarify changes in gene expression after 1) TNF-alpha + IFN-gamma, 2) TNF-alpha + IFN-gamma + terbutaline, and 3) TNF-alpha + IFN-gamma + GW9508 treatment. To this end, we set 4 groups of samples; 1) unstimulated group, 2) TNF-alpha + IFN-gamma-stimulated group, 3) TNF-alpha + IFN-gamma + terbutaline-stimulated group, and 4) TNF-alpha + IFN-gamma + GW9508-stimulated group. In each group, HaCaT cells were stimulated in triplicate wells (n=3).
Project description:total RNA from mouse (male c57BL/6) spleen labeled with Cy3 vs total RNA from mouse (male c57BL/6) B cells treated with Interferon-gamma (IFN gamma) labeled with Cy5- time course with repeats Keywords: ordered
Project description:Using a newly-developed workflow for quantitative newly synthesized proteome analysis (QuaNPA), featuring automated sample processing and multiplexed DIA (plexDIA) analysis, changes in the newly synthesized proteome of IFN-gamma treated Hela cells were monitored over time.
Project description:In this study, a novel high throughput method was successfully developed for the automated enrichment of newly synthesized proteins (NSPs) from complex environments, like cell culture media supplemented with fetal bovine serum (FBS). The proposed automated method allows for the robust and effective simultaneous enrichment of dozen of samples in a period of 3 hours. The enrichment method was comprehensively evaluated and validated, and was applied to the mass spectrometry (MS) analysis of the FBS-rich secretome of melanoma A375 cells treated with the immune cytokines interferon alpha (IFN-α) and gamma (IFN-γ).
Project description:Transcriptional activation of cultured mouse astrocytes in response to stimulation with CCM (complete cytokine mix: TNF-alpha, IL1-beta and IFN-gamma) at 4hr and 16hr time points.
Project description:Differential gene expression caused by 1h and 3h of IFN alpha or gamma treatment was analyzed in total cellular RNA of NIH-3T3 cells compared to mock; We used microarrays to analyze the effects of 1 and 3h of IFNalpha and gamma treatment in total cellular RNA Experiment Overall Design: NIH-3T3 cells ( 5th to 15th passage after thawing) were split from confluent plates 24h before start of the experiment. At the begin of the experiment about 80% confluency was reached. The experiment was started by applying fresh, prewarmed, CO2-equilibrated medium containing either mock, 100U/ml IFN alpha or 100 U/ml IFN gamma. Labeling was either started together with addition of IFN for 60 minutes or for 30 minutes starting 150 minutes after begin of treatment.
