Agilent g3 human gene expression 8 60 k arrays

RNA samples were analyzed for expression profiling using Agilent G3 Human Gene Expression 8 × 60 k Arrays (Agilent; Santa Clara, CA, USA). Total RNA quality for both microarray analyses was determined on an Agilent 2100 Bioanalyzer, with all samples having RNA integrity numbers greater than 8. Total RNA (250 ng) was used for labeling using a QuickAmp labeling kit (Agilent). Extracted RNA was labeled with cyanine (Cy)-3-CTP (PerkinElmer; Waltham, MA, USA) and reference RNA with (Cy)-5-CTP. Following purification of labeled cRNAs, 825 ng of Cy3- and Cy5-labeled cRNAs were combined and hybridized for 17 h at 65 °C in an Agilent hybridization oven. Microarrays were then washed and scanned using an Agilent DNA Microarray Scanner. The mRNA expression profiles in SAEC and HMVEC couture and monoculture are available in NCBI GEO with the accession number GSE129640.