GSM6199246: D6: control, replicate 6, RNAseq; Homo sapiens; RNA-Seq
Whole blood was collected from SCLS subjects during an asymptomatic interval or from age, sex, and ethnicity-matched healthy controls. Age-, sex-, and ethnicity-matched blood samples from anonymous donors were obtained from the NIH Blood Bank. We prepared buffy coat mononuclear cells from blood using Lymphocyte separation medium (MP Biomedicals). Cells were re-suspended in Endothelial Growth Medium (EGM-2, Lonza), and plated into culture wells coated with collagen I. The culture medium was changed every other day for several weeks until the colonies formed, and BOEC were expanded according to published protocols. RNA was extracted from BOECs using Direct-zol RNA Kit (Zymo Research). RNA libraries were prepared using the Illumina TruSeq® Stranded Total RNA Sample Preparation kit (Illumina) according to the manufacturer's instructions.