cell line: HepaRG cell line treatment: Treatment with DLM replicate: 3
Treatment protocol
The cells were individually exposed to 15 prototypical compounds for 24h and 72 hours at IC10 concentrations, i.e. 5 GTX (N-nitrosomorpholine, NMP, IC10=0.216mM; Hydroquinone, HQO, IC10=0.15mM; Hydrazine dihydrochloride, HHC, IC10=0.86mM; 2-acetylaminofluorene, TAF, IC10=0.0408mM; 2-amino-3-methylimidazo(4,5-f)quinoline, AMQ, IC10=0.003mM), 5 NGTX (Fumonisin B1, FMB, IC10=2.4mM; Cyclosporine A, CsA, IC10=0.0026mM; Acetamide, ACE, IC10=5.9; Diethylhexyl Phthalate, DHP, IC10=10mM; Ethanol, ETH, IC10=10mM), 5 NC (4-acetylaminofluorene, FAF, IC10=0.22; D,L-Menthol, DLM, IC10=1.2mM; Benzoin, BEN, IC10=0.345mM; Benzyl Alcohol, BEA, IC10=6.5mM; Triclosan, TRI, IC10=0.022mM).
Growth protocol
The human hepatoma-derived HepaRG cells (Biopredic International, France) were cultivated as previously described by A. Guillouzo et al. 168 (2007) 66-73. and P. Gripon et al 99 (2002) 15655-15660. At day 13 of cultivation, dimethylsulfoxide (DMSO)-containing medium (2%) was added for 7 days . At day 19 exposure with the selected compounds was initiated.
Extracted molecule
total RNA
Extraction protocol
Samples for RNA isolation were collected after 24 and 72 hours of exposure. Minimum three independent biological experiments were conducted for each compound. The total RNA extraction (RNA extraction kit, Qiagen), including a DNase digestion step, was done according to the manufacturer’s instructions.
Label
biotin
Label protocol
We used standard Affymetrix labeling protocols
Hybridization protocol
We used standard Affymetrix hybridisation rotocols
Evaluation of the robustness of classification of carcinogen-modified transcriptomic responses in HepaRG cells and the interlaboratory reproducibility of the model
