Thymocytes with iNKT rearrangements are found in the DP^dim population.

<p>A. FACS profile of thymocytes based on CD4 and CD8 expression and defined gates for DP^dim and DP^med thymocytes. B. FACS profile of thymocytes based on CD3 expression and defined gates for CD3^neg, CD3^med and CD3^hi. C. FACS profile of thymocytes based on CD4 and CD8 expression through CD3^neg and CD3^med populations. Numbers inside DP^dim gates represent the number of collected cells. D. Rearrangement analysis of Vα14-Jα18 in different sorted DP populations. The asterisk shows the population (CD3^med DP^dim) in which iNKT cells are observed, and lane “M” represents the iNKT rearrangement in the spleen of wild type C57BL/6 mouse and serves as size marker.</p

A model for the developmental dynamics of DN iNKT cells.

<p>The green box represents the thymus, and the red box the periphery. Filled ovals outlined in black identify cells transferred in the different experiments. Filled arrows and the timing next to them show the experimental findings. Open ovals represents specific compartments. Open arrows and the timing next to them are based on our interpretation. The gel insert shows the iNKT rearrangement in CD4⁺CD8^low and CD8⁺CD4^low cells.</p

iNKT rearrangement in DN thymocytes and spleen samples obtained from TCRα^−/− mice at different days post DP^dim injection.

a<p>Data from 7 experiments are summarized.</p

iNKT rearrangements in TCRα^−/− mice after intrathymic injection of DP^dim cells.

<p>The <i>X axis</i> shows days post injection, and the <i>Y axis</i> shows the percent of mice analyzed that had iNKT rearrangements. A. DN thymocytes were analyzed. The arrows and asterisks indicate statistically significant changes. Fisher’s exact test was used to compute the P value from contingency table (day 1& 3, P = 0.04; day 2&3, P = 0.03). B. Total splenocytes were analyzed. The correlation between the two linear regression lines was calculated based on the Zar’s method (P = 0.008). The actual data for each time point are in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043509#pone-0043509-t001" target="_blank">Table 1</a>.</p

DN iNKT cells can return to the thymus.

<p>DP^dim thymocytes were injected in the thymi of four TCR Cα^−/− mice and 6 days post transfer, the thymi were pooled and DN thymocytes were sorted. A.iNKT rearrangement from DN thymocytes 6 days post transfer of DP^dim. The newly generated DN day 6 thymocytes were transferred to the periphery of TCR Cα^−/− mice. B. iNKT rearrangements in the thymus at different days post peripheral transfer of day 6 DN thymocytes.</p