Affymetrix Generic Hybridization
At passage 7 of the HaCaT cells, cells plated in 12-well plates at 4.5e5 cells/well. A 100x stock of the chemical was freshly prepared and added to the corresponding wells, each exposure was performed in quadruplicate.
Samples were normalized to the average of 6 samples of the corresponding vehicle control samples, either ethanol or DMSO exposed cells. Then a log2 was applied to the data
The RNeasy kit (Qiagen) was used according to manufacturer instructions to obtain total RNA
The 3'IVT Epress labeling kit was used according to manufacturer protocol (Affymetrix)
HaCaT keratinocytes were cultured in T-flasks in DMEM medium supplemented with FCS, non-essential amino acids and penicillin/streptomicin. Around 80% confluence the cells were passaged.