RNA-Seq of CD3/CD28-stimulated healthy human peripheral blood CD4+ cells
CD4+ cells were isolated from fresh whole blood via positive magnetic bead selection (Miltenyi Biotec) and stimulated with anti-CD3/anti-CD28 antibody-coated beads (Invitrogen) for 4 hours at 37ºC. Lysates were prepared using QIAzol (QIAGEN) and stored at -80ºC for 3 to 8 months, and RNA was extracted using miRNAeasy (QIAGEN) immediately before cDNA library preparation and RNA-Seq.