array scanning and feature extraction protocol
P-AFFY-6 Affymetrix CEL analysis
Title: Affymetrix EukGE-WS2v4_450 Hybridization. Description:
Title: Affymetrix CHP Analysis (ExpressionStat). Description:
nucleic acid labeling protocol
P-AFFY-2 Affymetrix biotin labelling
Cultures were maintained in Dulbecco's modified Eagle's medium (DMEM;Invitrogen; Carlsbad, CA, USA) with L-glutamine and high glucose plus 10% FBS (Invitrogen) and 100U/mL penicillin streptomycin (Invitrogen) at 37oC and 5% CO 2. Cells were passaged at confluence by 0.25% trypsin EDTA (Invitrogen).
Total RNA was isolated from cells using RNeasy mini kit with columns following the manufacturer's protocol (Qiagen; Valencia, CA, USA). RNA integrity and purity were confirmed using aspectrophotometer and Agilent 2100 Bioanalyzer (Agilent Technologies; Palo Alto, CA, USA).
Freshly prepared synthetic human, porcine endothelin-1 (Et1) peptide (Sigma) was added to dermal fibroblast growth medium at 100nM for 24 hours.