Microarray Analysis of Whole Transcriptome

Total RNA was isolated by using the Nucleospin^® RNA kit protocol (Macherey–Nagel, Germany). Five hundred nanograms of total RNA was amplified using the Ambion total RNA amplification kit (Ambion, Carlsbad, CA), followed by hybridization of the synthesized cRNA with the Human HT-12 v4 Expression BeadChip kit (Illumina, San Diego, CA, USA), in accordance with the manufacturer’s protocol. Scanning of the chip was performed using Illumina BeadScan and BeadStudio (version 3.1). Data were processed with a package from Bioconductor (lumi) [23 (link), 24 (link)] using the free software environment R (http://www.r-project.org/). The microarray data has been registered in gene expression omnibus (https://www.ncbi.nlm.nih.gov/geo/) in NCBI (GSE95308).