NextSeq 500 paired end sequencing; ChIP sequencing on human, circulating, ex vivo isolated Naive CD4+ T cells and Naive Activated CD4+ T cells
ChIP sequencing on human, circulating, ex vivo isolated Naive CD4+ T cells and Naive Activated CD4+ T cells
CD4+ naive T cells have been isolated from peripheral PBMCs by density gradient centrifugation with Ficoll-Paque Plus. From PBMCs, T cells were negatively selected with magnetic separator (AutoMACS). CD4+ Naive were FACS sorted as CD4+CD25-CD127-/highCD45RO-. Activation to Th1 phenotype with IL-2, IL12 and anti-IL4 and stimulated with Dynabeads Human T-activator anti-CD3/anti-CD28 beads for 16 hours Crossilinked, sonicated and Immunoprecipitated DNA was purified with AMPure beads XP NEB Next Ultra 1
Experimental Factor: phenotype
CD4+CD25-CD127-/highCD45RO-
Experimental Factor: immunoprecipitate
anti-H3K4me3