D10 T cells were left untreated or pretreated with 10uM Akti1/2 for 1h then stimulated with biotinylated anti-mCD3/CD28 and streptavidin for 0, 2, 6 and 12hs
Growth protocol
The D10 Th2 T cell line was maintained in RPMI 1640, Supplemented with 10% heat-inactivated bovine growth serum (BGS; Hyclone), 2mM L-glutamine, 0.1mM nonessential amino acids, 50uM 2-ME, 100U/ml penicillin, 100ug/ml streptomycin and 25 IU/ml rhIL-2.
Extracted molecule
total RNA
Extraction protocol
RNA extraction was performed using a commercially available kit (RNeasy, Qiagen, Frederick,MD) according to the manufactures’ recommendations. RNA quality was confirmed based on a RNA integrity number >8 by use of an electrophoresis bioanalyzer (2100 Agilent Bioanalyser).
Label
biotin
Label protocol
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays
Hybridization protocol
Standard Illumina hybridization protocol
Scan protocol
Standard Illumina scanning protocol
Description
MouseRefSeq8 5773678020_F
Data processing
The data were normalised using cubic spline method
