SUPPLEMENTAL FIGURE 1. T cell development in T cell-specific RASA1deficient mice. All experiments were performed with thymi of littermate rasa1fl/fl and
rasa1fl/fl plck-cre mice. (A) Representative Western blot showing RASA1 expression in
whole thymi. (B) At left are shown representative two-color flow cytometry plots of
CD44 and CD25 expression among CD4-CD8- DN thymocytes. Numbers indicate the
percent of gated cells in each quadrant. Bar chart at right shows mean number + 1 SEM
of DN1 (CD44+CD25-), DN2 (CD44+CD25+), DN3 (CD44loCD25+) and DN4 (CD44CD25-) cells in thymi (rasa1fl/fl, n=7; rasa1fl/fl plck-cre, n=6). (C) Representative plots of
CD4 and CD8 expression (left) and bar chart (middle) showing mean number + 1 SEM of
DN, DP, CD4 SP and CD8 SP cells in thymi (rasa1fl/fl, n=10; rasa1fl/fl plck-cre, n=9). Bar
graphs at right depict mean ratios + 1 SEM of thymocyte subsets within genotypes. (D)
Representative flow cytometry plots of annexin V versus 7AAD staining upon the
indicated thymocyte populations (see Fig. 3 and Methods) from rasa1fl/fl and rasa1fl/fl
plck-cre mice. Similar results have been obtained in three repeat experiments. Statistical
significance was determined using the Student’s two-sample t-test.
SUPPLEMENTAL FIGURE 2. Influence of the plck-Cre transgene upon peripheral
T cell numbers. Two color flow cytometry plots depict CD44 and CD62L staining upon
splenic and LN CD4 and CD8 T cells from littermate rasa1+/+ and rasa1+/+ plck-cre
mice. Bar charts show mean number + 1 SEM of the indicated CD4 and CD8 populations
in LN (rasa1+/+, n=3; rasa1+/+ plck-cre, n=4). Statistical significance was determined
using the Student’s two-sample t-test.
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