DN1 thymocytes have the capacity to migrate to the LN. (A) Bulk thymocytes from Tcrβ/Tcrδ-double-knockout mice were stained for Lin, anti-CD44, and anti-CD25 and characterized by flow analysis. (B) CFSE labeled bulk thymocytes (4 × 106) from Tcrβ/Tcrδ-double knockout mice were injected intravenously into nude mice. A day after transplantation, spleen and LN cells were stained with Lin antibody cocktail, anti-CD25, and anti-CD44 and analyzed by flow cytometry. CFSE-positive donor-derived cells were gated (left), and the percentages of DN1 (CD44+CD25−), DN2 (CD44+CD25+), and DN3 (CD44−CD25+) cells were determined (right). (C) Total donor-derived cell number seeding the spleen and LN was calculated (right), followed by number and percentage of DN1 cells within this population (middle and left).
