Cell Culture, 100 mm dishes, minimum essential medium (Sigma) plus 10% fetal bovine serum (Life Technologies, Rockville, MD) and 2 mM glutamine in a 5% CO2 atmosphere.
Extracted molecule
total RNA
Extraction protocol
RNeasy® Mini Kit, according to the manufacturer's instructions.
Label
biotin
Label protocol
Biotinylated cRNA was prepared according to the standard Affymetrix protocol from 10 microg total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
Hybridization protocol
Following fragmentation, 15 microg of cRNA were mixed with spike controls and 2/3 was hybridized for 17 hr at 45C on GeneChip Human Genome 133 plus 2 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station per manufacturer's instructions.
Scan protocol
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000
Description
Gene expression data untreated HepG2 cells (control).
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 set to 1000.
