Gr-1+ (Ly6G+) cells were MACS-purified from splenocytes on day 7 after allogeneic (or syngeneic) BMT. Cells were used for RNA extraction when the purity was higher than 97%. For microarray hybridization, total RNA was isolated and purified using a DNA-free RNA isolation kit (RNAqueous-4PCR kit; Ambion, Austin, TX, USA).
Label
biotin
Label protocol
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays
Hybridization protocol
Amplified cRNA (1.5 Pg) was hybridized on Mouse WG-6 v.2 Expression BeadChip arrays (Illumina, San Diego, CA, USA), containing more than 45,281 well-annotated Ref transcripts.
Scan protocol
The array chips were scanned on a BeadArray reader (BeadStation 500G; Illumina)
Description
replicate 2 9482914090_E
Data processing
The raw microarray data were pre-processed through three steps: i) background correction was performed, and ii) the data were log-transformed to log2 scale and ჰ) normalized by the quantile normalization method implemented in Genome Studio software (Illumina).