genetic modification: cells stably expressing the oncogenes c-myc and Ha-ras under the control of the Tet-off regulatable promoter, konstitutive expression of the estradiol activatabel IRF-1hER fusion protein tissue: fibroblast
Treatment protocol
-
Growth protocol
DMEM, 10% FCS
Extracted molecule
total RNA
Extraction protocol
TRIZOL
Label
biotin
Label protocol
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 microg total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
Hybridization protocol
Following fragmentation, 10 microg of cRNA were hybridized for 16 hr at conditions recommended by the manufacturer. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
Scan protocol
GeneChips were scanned using the Affymetrix GCS 3000.
Description
Kröger et al., 2003. Oncogene 22:1045-1056
Data processing
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 150.
