GSM8723181: Effector Treg creERT2 control 4; Mus musculus; RNA-Seq
Single cell suspensions were prepared from pooled spleen and pLN, from an individual mouse of the indicated genotype. CD4+ cells were enriched by negative depletion using biotinylated anti-B220, anti-CD8 and anti-CD11b followed by Streptavidin Dynabeads (Dynal).Naïve or effector Treg and ex-Treg were sorted based on the indicated markers; all samples were sorted to >95% purity. RNA was prepared from sorted cells using the RNeasy Micro Plus kit (Qiagen), and quality was assessed using the Bioanalyzer RNA chip. cDNA was generated using the SMARTseq v4 ultra low input RNA kit for sequencing, and libraries prepared from the cDNA using the Nextera XT DNA library preparation kit.