Biosource strain/age/gender/disease state: neoplastically transformed cells derived from clonal subpopulations of derivation cell lines that were produced by 11 brief repeated exposures of normal diploid hepatic epithelial line WB-F344 cells to 5 ug/ml of N-methyl-N-Nitro-N-nitrosoguanidine
Growth protocol: Cells were routinely cultured in modified Eagle's medium (Life Technologies, Inc.) containing 10% FBS.
Treatment protocol: After growing to approximately 80% confluence, MEM culture medium was removed and replaced with a custom formulation of Chee's Essential Medium (CEM) containing 5% Fetal Bovine Serum with L-Arginine HCl at 0.168 gm/liter for an additional 18 hours.
Extraction protocol: Total RNA was isolated from cells after 18 hours of treatment using TRI reagent -- RNA isolation Reagent Method (TRI Reagent, RNA/DNA/Protein Isolation Reagent, Molecular Research Center Inc. Cincinnati, Ohio)
Labeling/Scanning/Hybrization/Quantification protocol: Target cRNA was prepared and hybridized (45 °C, 16 h) to GeneChip rat 230A arrays according to the manufacturer's directions (Affymetrix). Hybridized arrays were washed and stained using the GeneChip fluidics station 400 and scanned using the Agilent GeneArray scanner. Signals were analyzed using Microarray Suite 5.0 software (Affymetrix). Data obtained was analyzed with GeneSpring 6.0.
