Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of 3 genotypes in mouse

ABSTRACT: For transcription profiling six mouse T cells samples were analyzed: 3 genotypes 2 samples for each genotype were complared. Two wt animals-control;2 animals containing deficiency allele (Df11(1)) on chromosome 11 and 2 with the reciprocal duplication on chromosome 11(Dp11(1)). The part of the chromosome 11 deleted or duplicated is 0.8Mb between Gast and Hcd17b genes. Splenic CD4+ T cells were purified using anti-CD4 MiniMacs beads (Miltenyi Biotech, Bergisch Gladbach, Germany) according to the manufacturer's protocol and were plated at 106 cells/ml in complete RPMI 1640 medium (Sigma) supplemented with 10% fetal bovine serum (FBS), 0.3 mg/ml L-glutamine, 10 micromolar 2-mercaptoethanol, 100 U/ml penicillin, and 0.1 mg/ml streptomycin in 6 well plates coated overnight with anti-CD3 (5 _g/ml) and anti-CD28 (1 _g/ml) antibodies. After 5 days of culture cells were collected and prepared. Cells were suspended at 107 cells/ml in complete medium and cytokine secretion was induced with 10 ng/ml phorbol 12-myristate 13-acetate (PMA, Sigma) and 1 µg/ml ionomycin (Sigma). After 2 hours protein secretion was inhibited with brefeldin A (1:2000, eBioscience) and after 5 hours RNA from 2x106 cultured CD4+ T cells (RNAeasy kit, Qiagen, Hilden, Germany) was isolated following cytokine induction and 10 mg of RNA was used for hybridiation to mouse 430 2.0 array, Affymetrix, Santa Clara, CA arrays.

ORGANISM(S): Mus.Musculus

SUBMITTER: Olga Ermakova

PROVIDER: E-MTAB-447 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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