Lineage tracing of Ly49-expressing CD8 T-cells in the thymus

Multiple lines of evidence point to a thymic origin for Ly49+ CD8 T cells, however their development pathway remains elusive. We analyzed Ly49 receptors expression by thymocytes, after excluding unconventional T cells such as NKT, MAIT or gd T cells, by flow cytometry. Ly49+ thymocytes were then selected according to their expression of Ly49A, Ly49F or Ly49G2 receptors. Surface proteins and transcription factors expression was used to determine Ly49+ lineage differentiation in the thymus, using Slingshot.