Transcriptomic Analysis of Human Samples

Total RNA was extracted using the TRIzol method. RNA concentrations were determined with a NanoDrop 2000 spectrophotometer (Thermo Scientific, Pittsburgh, PA, USA). Total RNA from were screened for differentially expressed genes using an Agilent RNA 6000 Nano Kit (Agilent, Santa Clara, CA, USA), and a PrimeView Human GeneChip (Agilent, Santa Clara, CA, USA) was used for microarray analysis. RNA labelling and hybridisation to Agilent miRNA microarray chips were performed with a GeneChip Hybridization Wash and Stain Kit (Agilent, Santa Clara, CA, USA) (17 (link), 18 (link)). The GeneChip data was used to analyze by gene set enrichment analysis (GSEA), and differentially expressed genes were annotated using ingenuity pathway analysis (IPA) to predict path changes.