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Series GSE3296

Query DataSets for GSE3296

Status

Public on Oct 24, 2005

Title

Gene expression in MEFs in response to treatment with dipyridyl and trichostatin A

Organism

Mus musculus

Experiment type

Expression profiling by array

Summary

The CH1 protein interaction domain of the transcriptional coactivators p300 and CBP is thought to interact with HIF-1alpha and this interaction is thought to be critical to the expression of HIF-1alpha target genes in response to hypoxia. Trichostatin A (TSA), an inhibitor of histone deacetylases, has been reported to repress the expression of HIF-1alpha target genes. To test the requirement of the CH1 domain and TSA for gene expression in response to dipyridyl (a hypoxia mimetic), primary mouse embryonic fibroblasts (MEFs) were generated from C57Bl/6x129/Sv F2 e14.5 embryos that contain a deletion in the CH1 domain of three of four alleles of CBP and p300. The remaining allele of p300 or CBP was a conditional knock out allele. Control MEFs with only a single conditional knockout allele of p300 or CBP were also generated. At passage 3 MEFs were infected with Cre Adenovirus and grown until they had expanded at least 100 fold. Subconfluent MEFs were treated with ethanol vehicle or 100ng/ml TSA with 5% carbon dioxide at 37 C in a humid chamber for 30 min., followed by ethanol vehicle or 100 umdipyridyl (DP) for an additional 3hrs. Immediately after treatment, cells were lysed in Trizol for RNA extraction.
Keywords: genetic modification, dose response

Overall design

16 samples; 4 genotypes [CBP+/flox (flox1), p300 +/flox (flox2), CBP CH1/flox;p300 CH1/CH1 (triCH1flox1),CBP CH1/CH1;p300 CH1/flox (triCH1flox2)] , 4 treatments (EtOH, TSA, dipyridyl (DP) and TSA+DP).

Contributor(s)

Kasper LH, Boussouar F, Boyd K, Xu W, Biesen MA, Rehg J, Baudino TA, Cleveland JL, Brindle PK

Citation(s)

16237459

Submission date

Sep 12, 2005

Last update date

Feb 11, 2019

Contact name

Paul K. Brindle

E-mail(s)

paul.brindle@stjude.org