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Wnt & LIF signalling in mESC

ABSTRACT: The molecular processes underlying the properties of ESC are yet unknown even when it's well established that LIF/STAT3 is neccesary for the maintenance of pluripotency. Other pathways as Wnt are may be implicated in the regulation of the biological mechanisms in mESC. Work model: D3-ES cultivated with or without LIF and treated with chronic (7 days) low doses (50nM) of GSK3 β inhibitor (lithium). We used microarrays to detail the global profile of gene expression to identified different groups of up and down-regulated genes in the work model. mESC cultivated as it has been defined in the work model were prepared for RNA extraction and hybridization on Affymetrix microarrays. We analyzed 3 independent experiments per replicate, and we did three replicates per sample.

ORGANISM(S): Mus musculus

SUBMITTER: Patricia Ruiz-Ontañón

PROVIDER: E-GEOD-11628 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Wnt & LIF signalling in mESC

Project description:The molecular processes underlying the properties of ESC are yet unknown even when it's well established that LIF/STAT3 is neccesary for the maintenance of pluripotency. Other pathways as Wnt are may be implicated in the regulation of the biological mechanisms in mESC. Work model: D3-ES cultivated with or without LIF and treated with chronic (7 days) low doses (50nM) of GSK3 β inhibitor (lithium). We used microarrays to detail the global profile of gene expression to identified different groups of up and down-regulated genes in the work model.

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Establishment of a novel model to assess the function of proteins under in vivo conditions

Project description:We established a novel model to assess the function of proteins under in vivo conditions. The model relies on the expansion of HEK293 cells in immunodeficient NOD.Scid mice. To validate the novel model, we performed microarray gene expression profiling of NOD.Scid-expanded HEK293 cells relative to conventionally cultivated cells. Microarray analysis revealed that cell expansion in NOD.Scid mice restored an imbalanced chaperone system without inducing a major upregulation of the entire protein folding machinery. Human embryonic kidney (HEK293) cells were injected subcutaneously into immunodeficient NOD.Scid mice. After three weeks, the expanded cell pellet was isolated, and total RNA was extracted. In parallel, total RNA was prepared from HEK293 cells cultivated in vitro under standard cell culture conditions in a commercially available medium containing 450 mg/dl glucose (DMEM). Microarray gene expression profiling was performed to determine differential gene expression between in vivo expanded and in vitro cultivated HEK293 cells. Two biological replicates for each condition were made (in vitro cultivated HEK293 cells: Cells-1 and Cells-2; and NOD.Scid-expanded HEK293 cells: Scid-1 and Scid-2).

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Project description:Substantia nigra pars compacta (SNpc) is highly sensitive to normal aging and selectively degenerates in Parkinson's disease. However, ventral tegmental area (VTA), a region adjacent to SNpc, is less affected in PD. Until now, molecular mechanisms behind VTA aging have not been fully investigated using high throughput techniques. Here, aging-associated early changes in transcriptome of VTA were investigated comparing late middle-aged (18 months old) to young (2 months old) mice. Three age groups of C57 wild type mice were used in microarray analysis: young (2 months old), middle aged (10 months old), and late-middle aged (18 months old) mice. Four replicates were included in each age group and each replicate was pooled from 5 mice (5 mice/replicate x 4 replicates x 3 age groups). Total RNA was isolated from VTA for hybridization on Affymetrix microarrays.

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Age-mediated transcriptomic changes in adult mouse substantia nigra and ventral tegmental area

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2013-05-29 | E-GEOD-45045 | biostudies-arrayexpress

Expression data from Saccharomyces cerevisiae strains carrying the rna14-1 or the rna15-1 allele

Project description:In the yeast Saccharomyces cerevisiae, cleavage factor I (CFI) and cleavage and polyadenylation factor (CPF) build the core of the transcription termination machinery. CFI comprises the Rna14, Rna15, Pcf11, and Clp1 proteins, as well as the associated Hrp5 RNA-binding protein. We found that CFI participates in the DNA damage response and that rna14-1 shows synthetic growth defects with mutants of different repair pathways, including homologous recombination, non-homologous end joining, post replicative repair, mismatch repair, and nucleotide excision repair, implicating that impaired RNAPII termination and 3â-end processing decreases the cellular tolerance for DNA damage. Beyond replication progression defects, we found that bypass of the G1/S checkpoint in rna14-1 cells leads to synthetic sickness, accumulation of phosphorylated H2A, as well as increase in Rad52-foci and in recombination. Our data provide evidence that CFI dysfunction impairs RNAPII turnover, leading to replication hindrance and lower tolerance to exogenous DNA damage. These findings underscore the importance of coordination between transcription termination, DNA repair and replication in the maintenance of genomic stability. S. cerevisiae strains were grown in YPAD liquid culture at 30Â°C, total RNA was isolated and hybridized on Affymetrix microarrays.

2014-03-31 | E-GEOD-50947 | biostudies-arrayexpress

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