Fig. 3

E^BAB contributes to depletion of high affinity TCR clones in ex vivo TCR stimulation and the HY tg models. a Annexin V⁺Propidium Iodide (PI)⁻ fraction of total thymocytes stimulated with anti-CD3 and anti-CD28 antibodies (10 µg/ml) for 9 h. The percentage in mock sample (i.e., background) is subtracted. Data are pooled from 11 independent experiments (n = 3 WT–ΔE^BAB littermate pairs, n = 3 sex-matched WT–ΔE^BAB pairs, n = 1 WT–Bim^−/− littermate pair, n = 1 sex-matched WT–Bim^−/− pair, n = 3 sex-matched WT–Bim^−/−–ΔE^BAB trios, 5–20 weeks old). Edges of the box are the 25th and 75th percentiles, and error bars extend to the maximum and minimum. Outliers are defined as the data point that is located outside of q₃ + 1.5(q₃ − q₁) and q₁ − 1.5(q₃ − q₁), in which q₁ and q₃ are the 25th and 75th percentiles. b CD4 versus CD8 flow cytometric analysis of PI⁻HY-TCR⁺ thymocytes from female and male HY tg mice. The number in the plot is representative percentage of each gate. c−d DP thymocyte proportion (c) and cell numbers (d) of PI⁻HY-TCR⁺ thymocytes from female and male HY tg mice. e, f CD8 SP thymocyte proportion (e) and cell numbers (f) of PI⁻HY-TCR⁺ thymocytes from female and male HY tg mice. Data are representative of (b), or pooled from (c–f), six independent experiments (n = 1 Bim^+/−; HY⁺ and Bim^−/−; HY⁺ female mice, n = 5 Bim^+/−; HY⁺ and Bim^−/−; HY⁺ male mice, n = 3 E^BAB+/−; HY⁺ and ΔE^BAB; HY⁺ female mice, n = 5 E^BAB+/−; HY⁺ and ΔE^BAB; HY⁺ male mice, 6–8 weeks old). Each symbol in (c–f) represents an individual mouse; small horizontal lines indicate the means. n.s. not significant (P ≥ 0.05); *P < 0.05, **P < 0.01 (unpaired one-tailed Student’s t test or Mann–Whitney U test for Bim KO data, and unpaired two-tailed Student’s t test for ΔE^BAB data)