GSM2601498: TER119-SIINFEKL_1; Mus musculus; RNA-Seq
Mice were euthanized on day 4 and spleens were harvested for OTI CD8+ T cell isolation. Splenocytes were exposed for 5 min at RT to 0.155 M NH4Cl to lyse erythrocytes. A first round of isolation was performed for CD8+ T cell isolation using a CD8+ magnetic bead negative selection kit (Miltenyi Biotec), following the manufacturer’s instructions. The following staining steps were performed on ice. Cells were washed with PBS and stained for 15 min in PBS + 2% FBS with CD3e FITC, CD45.1 PE-Cy7, CD8 APC-e780, and CD4 Pacific blue antibodies (eBioscience). Cells were resuspended at a concentration of 8x10e6 cells/mL in PBS + 1 mM EDTA, and OTI CD8+ T cells were sorted using a FACS ARIAII cytometer. Viable transgenic OTI CD8+ T cells were gated based on their expression of the CD3e, CD8 and CD45.1 markers. Total RNA was extracted from sorted OTI CD8+ T cells using a RNeasy Plus Micro kit (Qiagen), following the manufacturer’s instructions. Libraries were generated using an Illumina RNA-SEQ kit (TruSeq® Stranded mRNA LT -- SetA, RS-122-2101).
GEO Accession
GSM2601498