RNA-Seq of Mus musculus: splenic OT-I CD8 T cells educated 2d with antigen-pulsed lymphatic endothelial cells
OT-I cells were selected by negative magnetic selection of CD8 cells from whole spleen of an OT-I mouse. They were then co-cultured on a monolayer of flow cytometry-sorted lymphatic endothelial cells (LECs; podoplanin- and CD31- double-positive), which had been pre-pulsed with ovalbumin and washed prior to addition of OT-I cells at a ~10:1 T:LEC ratio. The non-adherent OT-I cells were isolated and treated with Qiagen RNeasy Plus mini kit to isolate mRNA, which was then submitted to EPFL Gene Expression Core Facility for library preparation and sequencing.