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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Determination of HOXA1 and HOXA9 target genes with inducible HOX constructs

ABSTRACT: Target genes of HOXA1 and HOXA9 were determined with the help of inducible HOX-ER constructs. For this purpose bone marrow was harvested from C57BL/6 mice and c-Kit positive progenitors were enriched by magnetic sorting (MACS technology, Miltenyi, Germany). After retroviral infection by spinoculation the cells were plated for three rounds in methylcellulose. Subsequently cells were maintained in liquid culture in RPMI1640 supplemented with murine recombinant cytokines at 100ng/ml (stem cell factor) and 10ng/ml (IL-3, IL-6, granulocyte/macrophage-colony stimulating factor) as well as 100nM 4-hydroxy-tamoxifen (TAM) for HOX activation.

For microarray analysis 4 independent cell lines were created for each HOX construct. RNA was isolated from cells in the presence of TAM and 72h after withdrawal of the inductor. Pairs of RNA replicates were pooled and two +TAM as well as two -TAM duplicates per HOX gene were hybridized to Agilent44K mouse expression arrays. Array processing and evaluation was done commercially according to Agilent specifications by ImaGenes (now SourceBioScience), Berlin, Germany.

ORGANISM(S): Mus musculus

SUBMITTER: Robert Slany

PROVIDER: E-MEXP-3648 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

HOX genes regulate Rac1 activity in hematopoietic cells through control of Vav2 expression.

Breitinger C C Maethner E E Garcia-Cuellar M-P MP Schambony A A Fischer K D KD Schilling K K Slany R K RK

Leukemia 20120620 1

PMID: 22713647

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