Human mononuclear cells were isolated by Ficoll-Paque centrifugation in a discontinuous gradient from buffy coats from healthy blood donors. Cells were differentiated in macrophage-SFM medium supplemented with 70 U/ml human granulocyte macrophage colony-stimulating factor. After 3 days, the medium was changed to macrophage-SFM without GM-CSF and cells were cultured up to 8 days
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated with the RNeasy kit (Qiagen).
Label
PE
Description
Total RNA was isolated with the RNeasy kit (Qiagen) from macrophages incubated at normoxia (21% oxygen) or hypoxia (0% oxygen) for 24 h. Two μg RNA from 4 donors incubated at normoxia and 4 donors at hypoxia was pooled respectively and these 2 pools were used for the target preparation and analyzed on duplicate DNA microarrays (Hu95A; Affymetrix).
Data processing
MAS 5, signal algorithm, global scaling (target intensity 500)
This is the final calculated measurement (MAS 5 signal) for each probe set identifier that has been made comparable, through scaling (target intensity 500), across all samples and rows
ABS_CALL
Presence or Absence of gene transcript in sample (MAS 5 detection call)
