Transcriptomics

Dataset Information

24

Rapamycin effect on Wild Type and TSC deficient Mouse Embryonic Fibroblasts : Time Course


ABSTRACT: Aberrant activation of the mammalian target of rapamycin (mTOR) complex 1 (mTORC1) is a common molecular event in a large variety of pathological settings, including genetic tumor syndromes, cancer, and obesity. However, the cell intrinsic consequences of mTORC1 activation remain poorly defined. Here, we identify global trancriptional changes in TSC1 and TSC2 null MEFs, which exhibit constitutive activation of mTORC1, compared to wild-type littermate control lines. A rapamycin time course is included to determine those changes that are dependent on mTORC1 signaling, revealing mTORC1 induced and repressed transcripts. In order to identify mTORC1-dependent transcriptional changes, we compared wild-type MEFs to both Tsc1-/- and Tsc2-/- MEFs following serum starvation, where mTORC1 signaling is off in wild-type cells and fully active in TSC-deficient cells. All cell lines were serum-starved for 24 h, and the Tsc1-/- and Tsc2-/- cells were treated with a time course of rapamycin prior to the isolation of mRNA for microarray analysis. Overall design: Immortalized wild-type (Tsc2+/+ p53-/-), Tsc1-/- (p53+/+, 3T3-immortalized), and Tsc2-/- (p53-/-, derived from a littermate of the wild-type cell line) MEFs are the three cell lines used in this study and were derived in the laboratory of David J. Kwiatkowski (Brigham and Women's Hospital, Harvard Medical School, Boston, MA). Wild-type and null MEFs were grown to 70% confluence in 10 cm plates and were serum starved for 24 h in the presence of vehicle (DMSO) for 24 h or rapamycin (20 nM) for 2, 6, 12, or 24 h. All vehicle-treated samples (0 h time points) were plated in triplicate and all rapamycin time course samples were plated in duplicate. For each replicate, expression analysis was performed by hybridization to an Affymetrix Mouse 430_2 oligonucleotide microarray chip.

INSTRUMENT(S): [Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array

ORGANISM(S): Mus musculus  

SUBMITTER: Pichai Raman  

PROVIDER: GSE21755 | GEO | 2010-07-30

SECONDARY ACCESSION(S): PRJNA127325

REPOSITORIES: GEO

Similar Datasets

2010-07-30 | E-GEOD-21755 | ArrayExpress
2011-08-03 | GSE27982 | GEO
2011-08-02 | E-GEOD-28021 | ArrayExpress
2011-08-03 | GSE28021 | GEO
2011-08-02 | E-GEOD-27982 | ArrayExpress
2015-08-12 | BIOMD0000000581 | BioModels
| GSE16944 | GEO
2018-10-24 | PXD002670 | Pride
2016-05-04 | PXD001340 | Pride
2011-11-12 | E-GEOD-25331 | ArrayExpress