Transcription profiling of forskolin-synchronized NIH3T3 cells to detect rhythmic transcripts
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Michael Hughes
PROVIDER: E-GEOD-11922 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Forskolin treatment induces activation of protein kinase A. This activation protects transformed cells from death induced by glucose starvation. We used microarray to identify the mechanisms involved in such a pro-survival effect. NIH3T3 and NIH3T3-K-ras were plated at 1mM glucose and then daily treated or not with 10uM FSK until 72h of growth. The microarray analysis has been performed at 72h in order to identify specific mechanisms associate with transformed cells survival.
2015-10-24 | E-GEOD-68266 | biostudies-arrayexpress
Project description:Regulation of organ size is important for development and tissue homeostasis. In Drosophila, Hippo signaling controls organ size by regulating the activity of a TEAD transcription factor, Scalloped, through modulation of its coactivator protein Yki. The role of mammalian Tead proteins in growth regulation, however, remains unknown. Here we examined the role of mouse Tead proteins in growth regulation. In NIH3T3 cells, cell density and Hippo signaling regulated the activity of Tead proteins by modulating nuclear localization of a Yki homologue, Yap, and the resulting change in Tead activity altered cell proliferation. Tead2-VP16 mimicked Yap overexpression, including increased cell proliferation, reduced cell death, promotion of EMT, lack of cell contact inhibition, and promotion of tumor formation. Growth promoting activities of various Yap mutants correlated with their Tead-coactivator activities. Tead2-VP16 and Yap regulated largely overlapping sets of genes. However, only a few of the Tead/Yapregulated genes in NIH3T3 cells were affected in Tead1-/-;Tead2-/- or Yap-/- embryos. Most of the previously identified Yap-regulated genes were not affected in NIH3T3 cells or mutant mice. In embryos, levels of nuclear Yap and Tead1 varied depending on cell types. Strong nuclear accumulation of Yap and Tead1 were seen in myocardium, correlating with requirements of Tead1 for proliferation. However, their distribution did not always correlate with proliferation. Taken together, mammalian Tead proteins regulate cell proliferation and contact inhibition as a transcriptional mediator of Hippo signaling, but the mechanisms by which Tead/Yap regulate cell proliferation differ depending on cell types, and Tead, Yap and Hippo signaling may play multiple roles in mouse embryos. We used microarrays to know the gene expression profiles regurated by Tead2-VP16, Tead2-EnR, Yap, and cell density in NIH3T3 cells. Keywords: Cell density, genetic modification Tead2-VP16-, Tead2-EnR-, Yap- and control vector-expressing cells were cultured at low or high density for RNA extraction and hybridization on Affymetrix microarrays.
2008-11-20 | E-GEOD-12498 | biostudies-arrayexpress
Project description:NIH3T3 in the middle of G0 to G1 transion consists of the cells which is still staying G0 phase and the cells which enters G1. Monitoring the expressions of p27 and Cdt1 enables to distinguish these two; p27+/Cdt1+ cells as the cells in G0 phase and p27-Cdt1+ cells as G1 phase We sorted p27+Cdt1+ (G0) NIH3T3 cells and p27-Cdt1+ (G1) NIH3T3 cells in the middle of G0-G1 transition, 5 hours after serum addition following the serum addition using mVenus-p27K- and mCherry-hCdt1(30/120) . We compared the expression profiles of these NIH3T3 cells in G0 and G1 phases and identified the genes upregulated in G0 or G1 phases. The p27+Cdt1+ (G0) NIH3T3 cells and p27-Cdt1+ (G1) NIH3T3 cells were sorted by FACS for RNA extraction and hybridization on Affymetrix microarrays.
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Project description:Expression profiling of normal NIH3T3 and transformed NIH3T3 K-ras cell lines grown for 72 hours in optimal glucose availability (25 mM glucose) or low glucose availability (1 mM). Low glucose induces apoptosis in transformed cells as compared to normal ones. We performed genome-wide analysis by using Affymetrix GeneChip oligonucleotide microarrays to identify those genes whose expression levels are modulated by glucose availability in NIH3T3 and NIH3T3 K-ras cell lines. The cells, to be used for the transcription analysis, were collected at 18h from the initial seeding, time corresponding to the change of medium (25 or 1 mM glucose, indicated as T0) and then at 24, 48 and 72h upon medium change.
2011-07-13 | E-GEOD-29962 | biostudies-arrayexpress
Project description:Identification of cyclical expressed coding and non-coding genes during the circadian rhythm in NIH3T3 cells. NIH3T3 cells were synchronized for their circadian rhythm and RNA sequencing were performed at several time points along the rhythm. This data was used to identify cyclical expressed genes as well as long intergenic non-coding RNAs. NIH3T3 cells were synchronized with 100 nM Dexamethasone for 2 hours, then medium was changed to normal culture medium (0h). Every 4 hours cells were harvested, RNA isolated and RNAseq performed.
2015-12-31 | E-GEOD-66243 | biostudies-arrayexpress
Project description:Fibroblasts usually mediate acute wound healing and long-term tissue remodeling with scarring in tissue injury. In myocardial infarction (MI), following a prolonged lack of oxygen supply, necrotized cardiomyocytes become replaced by secreted extracellular matrix proteins produced by fibroblasts. Dendritic cells (DCs) act as inflammatory cells and can migrate from the bone marrow to the infarct areas and infarct border areas to mediate collagen accumulation after MI, whereas trichostatin A (TSA) can regulate the apoptosis and proliferation of the fibroblasts and affect DCs functions under oxygen–glucose deprivation (OGD) conditions. In this study, we used proteomics to investigate the effects of TSA and bone marrow-derived dendritic cells (BMDCs) on NIH3T3 fibroblasts under OGD conditions. Results showed that the fatty acid degradation pathway was significantly upregulated in NIH3T3 cells under OGD conditions, and the fatty acid synthesis pathway was significantly downregulated in NIH3T3 cells treated with BMDCs conditioned media with TSA (BMDCs-CM[TSA]) under OGD conditions. Meanwhile, the BMDCs-CM(TSA) significantly decreased the levels of triglycerides and free fatty acids and mediated ten fatty acid metabolism-related proteins in the NIH3T3 cells under OGD conditions. Summarily, the proteomic analysis showed that TSA and BMDCs affect fatty acid metabolism in NIH3T3 cells under OGD conditions.
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Project description:Forskolin treatment induces activation of protein kinase A. This activation protects transformed cells from death induced by glucose starvation. We used microarray to identify the mechanisms involved in such a pro-survival effect.
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Project description:To investigate biological function of MARVELD1, we have employed whole genome microarray expression profiling as a platform to analyze gene expression alternation influenced by MARVELD1. MARVELD1 stably transfected NIH3T3 and control vector cells were used to compare their gene expression profiling. Total RNA from MARVELD1 overexpressing NIH3T3 cells and control cells were extracted to analyze their gene expression profiles by using mouse whole genome expression microarray.
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2020-10-28 | PXD019977 | Pride
Project description:UBF is an essential RNA Polymerase I (Pol I)-transcription factor. Our research investigates extra roles for UBF in regulation of Pol II gene expression. Our gene expression data identifies genes that are differentially regulated following UBF knockdown by siRNA. RNA was extracted from NIH3T3 cells after 48 hour of transfection with two independent siRNA oligonucleotides targeting UBF, sirEGFP or Mock control. A total of 12 samples were analysed using Affymetrix arrays.We include the expression data from three biological replicate experiments
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