Monocytes from cryopreserved PBMC of normal and CGVHD patients were enriched by gating on FSChi SSChi CD3– CD4dim cells using an Influx Cell Sorter (Becton Dickinson Immunocytometry Systems)
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from monocytes by using RNeasy kit (Qiagen, Germantown, , MD, USA)
Label
Biotin
Label protocol
300ng of total RNA per sample was amplified according to manufacturer's instructions (Ambion WT Expression Kit). Next cDNAs were fragmented, biotinylated, and hybridized to the GeneChip Human Gene 1.0 ST Arrays (Affymetrix WT Terminal Labeling Kit).
Hybridization protocol
300ng of total RNA per sample was amplified according to manufacturer's instructions (Ambion WT Expression Kit). Next cDNAs were fragmented, biotinylated, and hybridized to the GeneChip Human Gene 1.0 ST Arrays (Affymetrix WT Terminal Labeling Kit).
Scan protocol
scanning was carried out with the GeneChip Scanner 3000 and image analysis with the Affymetrix GeneChip Command Console Scan Control(all from Affymetrix, Santa Clara, CA).
Description
gene expression data
Data processing
Expression data were normalized, background-corrected, and summarized using the RNA algorithm
