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. 2019 Jul 19;216(10):2427–2447. doi: 10.1084/jem.20181959

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© 2019 Martínez-Riaño et al.

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Figure 1. — Reduced percentage of DP thymocytes in Rras2-deficient mice. (A) Flow cytometry analysis of WT and Rras2-deficient thymocytes in 6-wk-old C57BL/6 mice according to the expression of CD4 and CD8α markers. Bar plots to the right represent the number of CD4ˉCD8ˉ (DN), CD4⁺CD8⁺ (DP), CD4⁺CD8ˉ (CD4SP), CD4ˉCD8⁺ (CD8SP), and total thymocytes in both genotypes (n = 8 mice per group). Blue circles, Rras2^+/+; red squares, Rras2⁻^/−. (B) Percentages of DN, DP, SP4, and DP8 thymocytes in mice analyzed as in A. n = 11 mice per group. (C) Flow cytometry analysis of CD3 expression in the indicated thymocyte populations. An overlay plot of Rras2^+/+ (continuous line) and Rras2⁻^/− (broken lines) is shown on the left, and quantification of CD3 expression is shown in the bar blots on the right. n = 4 mice per group. MFI, mean fluorescence intensity. (D) Number of DP thymocytes according to the expression of CD3. Two-color plots show CD3 expression versus DP thymocyte size. Bar plots below represent the number of DP thymocytes expressing or not expressing CD3 for both genotypes Rras2^+/+ (blue circles) and Rras2⁻^/− (red squares). n = 9 mice per group. FS, forward scatter. (E) Flow cytometry analysis of WT and Rras2-deficient thymocytes in AND^Tg mice in a pure b/b background and in a mixed k/b background after gating within the transgenic Vβ3^high thymocytes. Bar plots to the right represent the percentage of thymocyte subpopulations for both genotypes. Blue circles, Rras2^+/+; red squares, Rras2⁻^/−. n = 4–7 mice per group. (F) Number of DN, DP, SP4, and DP8 thymocytes in AND TCR mice in a pure b/b background and in a mixed k/b background. Blue circles, Rras2^+/+; red squares, Rras2⁻^/−. n = 4–6 mice per group. Quantitative data in all panels are means ± SEM. *, P < 0.05; **, P < 0.005; ****, P < 0.0005; n.s., not significant (unpaired two-tailed Student’s t test). All phenotyping experiments in this figure were repeated at least three times.