Mus musculus Phenotype or Genotype

we separated naive CD8 or naive CD4 cells from spleen cells of C57BL/6 mice,then ex vivo stimulated naive cells with TGF-ß, IL-2 and anti-CD3/CD28 beads, then wegot CD4+ iTreg cells or CD8+ iTreg cells. We also directly separated nTreg (CD4+CD25+) from spleen cells. The goal of the study is to find out the different phenotype and marker of CD8+CD103+ iTreg, CD4+ iTreg and nTreg. This three kind of Treg all showed potent therapeutic effect on autoimmune diseases according to many studies.