Exposures of 24 hours in duration were initiated once flasks were 90 ± 10% confluent using the test chemicals NiCl2, CdCl2, and Na2Cr2O7.
Growth protocol
H4-II-E-C3 cells (ATCC, Manassas, VA) were grown in Dulbecco's Modified Eagle's Medium (DMEM; Lonza,Walkersville, MD) containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA) and 10 mL Glutamax (Invitrogen) in T75 flasks incubated at 37 °C with 5% carbon dioxide.
Extracted molecule
total RNA
Extraction protocol
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
Label
biotin
Label protocol
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 7.5 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
Hybridization protocol
Samples were hybridized on the GeneChip Rat Genome 230 2.0 Array using Affymetrix instrumentation according to the manufacturer’s instructions
Scan protocol
Affymetrix protocol
Data processing
Microarray data was processed for background adjustment, normalization, and summarization using the Robust Multi-Array Averaging method (RMA) using Partek Genomic Suite (GS) software (Version 6.4 Copyright 2009, St. Louis, MO)
