GSM7991797: VAS90; Homo sapiens; RNA-Seq
Peripheral blood mononuclear cells were acquired from whole blood by density gradient centrifugation using Ficoll-Paque (Rafer, Zaragoza, Spain). These cells were then incubated with CD4-APC conjugated antibodies (MiltenyiBiotec, Germany) in staining buffer (PBS with 2mM of EDTA and 4% FBS) for 20 minutes. Next, flow cytometry sorting was used to isolate CD4+ T cells through positive selection. Finally, both genomic DNA and total RNA were extracted from the same cell pellet by using AllPrep DNA/RNA/miRNA Universal kit (Qiagen, Hilden, Germany).  Total RNA quality was first determined by the 2100 Bioanalyzer System (Agilent, CA, USA), 1 ug of those samples with excellent quality RNA (RNA integrity number >7) was used for library synthesis with the TruSeq Stranded mRNA Library Prep Kit (Illumina, CA, USA) according to the manufacturer's protocol