title: Sample 2
organism: Mus musculus
description: Protocols: Foxp3-GFP KI mice were infected with 200 pfu of the Lymphocytic choriomeningitis virus (strain WE) (LCMV WE strain) or left uninfected (naive). Infection with low dose LCMV WE induces an acute systemic Th1-driven immune response which is accompanied by a peak in CD4+Foxp3+CXCR3+ expressing Treg cells at day 14 post infection. At day 14 post infection spleens and peripheral LNs were collected, mashed through a metal grid or a 70 µm cell strainer, respectively to obtain a single cell suspension. Red blood cells were lysed via ACK lysis. CD4+ T cells were MACS sorted using anti-CD4 beads (Miltenyi) prior to flow sorting of CD4+Foxp3-GFP+ Treg cells from naive mice or CD4+Foxp3-GFP+CXCR3+ Treg cells from LCMV-infected mice. RNA was extracted using the Qiagen RNeasy Micro Kit. Libraries for sequencing were generated with the TruSeq RNA Sample Preparation kit (Illumina).