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. 2007 Feb;37(2):487–500. doi: 10.1002/eji.200636503

Figure 1.

Figure 1

Analysis of DN3 and DN4 foetal and adult wild type thymocytes. (A) Gates for the identification of DN3 and DN4 subsets (shown for E16.5 thymus). Thymocytes in the live FSC/SSC gate (left) were determined as DN3 or DN4 when falling in the gate negative for anti-CD3/CD4/CD8/CD44 and positive for anti-Thy1.2 antibody staining (middle) and positive or negative for anti-CD25 staining (right), respectively. (B–D) Percentages of icTCRβ+ cells among CD44–CD25+ TN (DN3) and CD44–CD25– TN (DN4) subsets are shown for E15.5 (B), E16.5 (C) and adult thymus (D). The percentage of icTCRγδ+ cells in the icTCRβ– DN4 subset on E16.5 is also shown (C). (E) Expression of icCD3 in DN3, icTCRβ– and icTCRβ+ DN4 thymocytes. Expression of icCD3 in DN3 cells is shown in the top, icTCRβ– DN4 in the middle and icTCRβ+ DN4 cells in the lower panel. Percentages of high and low icCD3 expression are shown for the DN3 and icTCRβ– DN4 subsets. The dotted line in all histograms represents the expression of icCD3 in B lymphocytes, similar in size to thymocytes and isolated from lymph nodes of adult wild-type mice, and is a negative control for the icCD3 staining. (F) Kinetics of icTCRβ– and icTCRβ+ DN4 thymocyte number. The number of icTCRβ– DN4 and icTCRβ+ DN4 thymocytes are shown for E15.5, E16.5 and adult thymi. Bars represent the mean of at least three thymi and error bars the standard error of the mean.