Primary keratinocytes were cultured in Epilife medium (Cascade Biologics, Portland, OR, U.S.A.) with EDGS (Cascade Biologic) and Penicillin Streptomycin (100 IU mL-1 and 0.1 mg mL-1, respectively) (Gibco, Carlsbad, CA, U.S.A.) at 37 °C. Once 80% confluent, the medium was supplemented with or without the following cytokines: recombinant human (rh)-IL-17 (R&D System, Minneapolis, MN, U.S.A.) 200 ng mL-1 and TNF-α (Sigma-Aldrich Corp, St. Louis, MO, USA) (1 ng mL-1 or 10 ng mL-1) for 24 h before harvesting for other analyses.
Extracted molecule
total RNA
Extraction protocol
Qiagen RNeasy Kit protocol
Label
biotin
Label protocol
GeneChip Affymetrix protocol
Hybridization protocol
GeneChip Affymetrix protocol
Scan protocol
Hewlett–Packard Gene Array Scanner
Data processing
GCRMA (using gcrma package from R/Bioconductor). Each platform was preprocess separately, then merged. An extra quantile normalization was needed to normalized across platforms.
