[HT_HG-U133_Plus_PM] Affymetrix Human Genome U133 Plus PM Array [Brainarray Version 13, HTHGU133PlusPM_Hs_ENTREZG]

Applicability of a keratinocyte gene signature to predict skin sensitizing potential

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HaCaT human keratinocytes were exposed for 4 hours to 26 compounds at a concentration  that resulted in 20% cell death after 24 hours of exposure.

Compound

Dose

Human keratinocyte sensitizing potential

Generation and validation of a gene signature predictive for skin sensitizing potential in human keratinocytes

HaCaT human keratinocytes were exposed for 4 hours to 26 compounds at a concentration  that resulted in 20% cell death after 24 hours of exposure.


Affymetrix:Protocol:Hybridization-Unknown

P-MTAB-24950

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. 

P-MTAB-24951

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. 

P-MTAB-24952

The RNeasy kit (Qiagen) was used according to manufacturer instructions to obtain total RNA

P-MTAB-24953

The 3'IVT Epress labeling kit was used according to manufacturer protocol (Affymetrix)

P-MTAB-24954

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

Human keratinocyte sensitizing potential

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Samples