Human genome u219 96 genetitan array

Microarray profiling was performed on 140 untreated or laser-treated samples using the HG-U219 array (Affymetrix) as similarly described for the aged dermis samples [8 (link)]. Flash-frozen, bisected 4 mm full-thickness skin punch biopsies were homogenized in TRIzol reagent (ThermoFisher, Waltham, MA) and total RNA was extracted according to the manufacturer’s protocol. RNA was further purified using RNEasy spin columns (QIAGEN, Hilden, Germany). Biotinylated cRNA libraries were synthesized from 500 ng of total RNA using the Affymetrix HT 3’ IVT Express kit (Affymetrix, Santa Clara, CA) and the Beckman Biomek FXp Laboratory Automation Workstation (Beckman Coulter, Brea, CA). Biotinylated cRNA was subsequently fragmented by limited alkaline hydrolysis and then hybridized overnight to the Affymetrix Human Genome U219-96 GeneTitan array. Plates were scanned using the Affymetrix GeneTitan Instrument utilizing AGCC GeneTitan Instrument Control software version 4.2.0.1566. Image data was summarized using Affymetrix PLIER algorithm with quantile normalization. Microarray data from this study as well as the aged dermis study have been deposited in the Gene Expression Omnibus (GEO) under GSE168760 and GSE139300, respectively.