NIH-3T3 cells were pretreated for 15 min with either DMSO (mock) or cycloheximide followed by addition of either mock, 100 U/ml IFNalpha or 100 U/ml IFNgamma for 1h. During the last 30 min, 500 µM 4-thiouridine was added to cell culture medium.
Extracted molecule
total RNA
Extraction protocol
Total cellular RNA was isolated using Trizol reagent and nascent RNA was purified as described (Dölken et al. RNA 2008) .
Label
biotin
Label protocol
standard Affy
Hybridization protocol
standard Affy
Scan protocol
AGCC
Description
D4_CHX_Mock_RepD nascent RNA, 30-60 min gene expression data