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. 2012 Jul 30;209(8):1409–1417. doi: 10.1084/jem.20120846

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© 2012 Martins et al.

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Figure 3. — Intrathymic differentiation in the absence of continuous progenitor import. (A and B) 9 wk after thymus transplantation, Rag2^−/−γ_c^−/−Kit^W/Wv recipients and C57BL/6 control mice received 1 mg BrdU and thymocytes were analyzed 2 h or 5 d later. Dot plots in A show expression of CD4 and CD8 by BrdU⁺ cells and represent examples of individual thymus grafts. Numbers indicate percentages of cells in each region. (B) Percentages of BrdU⁺ cells within each indicated population (DN, DP, CD4, and CD8) were analyzed 2 h and 5 d after BrdU injection. Results from one experiment are shown, where each dot is an individual C57BL/6 thymus (B6) or an individual thymus graft (G). The 2-h pulse labeling was done in 3 independent experiments, in which 3, 4, or 8 grafts with DP were analyzed. The 5-d chase experiment was performed once, and 9 grafts with DP were analyzed. (C) B6/Ly5.1 control thymus, and a representative graft after 10 wk in a Rag2^−/−γ_c^−/−Kit^W/Wv host, were analyzed for the indicated markers. Cells stained for CD44 and CD25 were pregated as Lin⁻CD45.1⁺ donor cells, and cells stained for CD44 and Kit were pregated as Lin⁻CD25⁻CD45.1⁺ donor cells. Numbers indicate percentages of cells in each gate. A total of 3 independent experiments were performed, in each of which 3–10 grafts with cellularities >10⁶ cells were analyzed.