Raw unaligned whole-blood RNAseq data
Total RNA was extracted from whole blood samples in PAXgene blood RNA tubes using PAXgene Blood RNA Kits (Qiagen, Germantown, MD, USA) in accordance with the manufacturer's protocol (Azenta Life Sciences, South Plainfield, NJ, USA). Total RNA samples were quantified using a Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked with a 4200 TapeStation (Agilent Technologies, Palo Alto, CA, USA). Samples were initially treated with TURBO DNase (Thermo Fisher Scientific, Waltham, MA, USA) to remove DNA contaminants, after which rRNA and globin depletion were performed using QIAseq® FastSelect™−rRNA HMR and −Globin kits (Qiagen). RNA sequencing libraries were constructed using the NEBNext Ultra II RNA Library Preparation Kit for Illumina (New England Biolabs, Inc., Ipswich MA, USA). Sequencing libraries were validated using the Agilent Tapestation 4200 (Agilent Technologies), and quantified using a Qubit 2.0 Fluorometer (ThermoFisher Scientific) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). Sequencing libraries were multiplexed, clustered on nine lanes of a flowcell, and loaded on the Illumina HiSeq 4000 instrument according to the manufacturer's instructions (Illumina, Inc., San Diego, CA, USA). Samples were sequenced using a 2x150 paired-end configuration. Raw sequence data were converted into FASTQ files and de-multiplexed using Illumina's bcl2fastq 2.17 software.