incubation time: 4 treatment: Interferon-alpha-2a treatment dose u/ml: 100 ifna responder: + tgfb responder: ++ ifna and tgfb responder: +++ culture vessel: T25 flask timepoint 4h treatment: not same day ivt on extraction day: FALSE sentrix barcode: 1461896068 sample section: H cell line: ME15
Treatment protocol
The cell lines were grown without (control) or with 100 U/ml human IFN- 2A (Roferon, F. Hoffmann-La Roche Ltd., Basel, Switzerland) for 4 or 24 h.
Growth protocol
Cell lines were cultured at 37°C in a 5% CO2 atmosphere in RPMI 1640 medium (GIBCO Life Sciences, Paisley, U.K.) supplemented with 10% fetal bovine serum (FBS), L-glutamine (2 mM), sodium pyruvate (1 mM), nonessential amino acids, antibiotics, and 10 mM HEPES buffer.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted with TRI reagent (Molecular Research Center, Inc., Cincinnati, OH).
Label
Cy3
Label protocol
TotalRNA was biotin-labeled using a commercial kit and the protocol supplied (Agilent Technologies, Basel, Switzerland).
Hybridization protocol
Labeled, unfragmented cRNA (500 ng) of each cell line was hybridized to commercial microarrays using the manufacturer’s protocol (Sentrix HumanRef-8 Expression BeadChips; Illumina, Inc., San Diego, CA).
Scan protocol
Data were collected using a confocal scanner (Illumina beadstation).
Description
Tissue_Endothelial
Data processing
Data analysis was performed using Illumina Genomestudio (www.illumina.com) and R/Bioconductor software. The data was normalized using the package limma and the functions lumiT and lumiN(method='quantile).
