Figure 4: Pre-rearranged TCR corrected the T-Red phenotype.

(a,b) Histograms of CD44 levels (a) and percentages of the CD44 high memory/activated T-cell phenotype (b) in peripheral blood of P14, OT-I and OT-II TCR transgenic mice of T-Red- or WT backgrounds. Mice up to 12 weeks old were used. Representative FACS plots from more than three independent experiments are shown. One-way ANOVA with post hoc Dunnett’s test was used in b. (c) Total number of splenocytes and CD8+Vβ8.1/8.2+ T cells in P14 TCR transgenic mice (Vβ8.1+) of T-Red or WT backgrounds (6–8 weeks old). (d) Absolute cell numbers of thymocyte subpopulations in OT-I and T-Red/OT-I mice (8–9 weeks old). (e,f) Bone marrow cells from WT (CD45.2/CD90.1), T-Red (CD45.1/CD45.2/CD90.2), OT-I (CD45.2/CD90.1/CD90.2) and T-Red/OT-I (CD45.2/CD90.2) mice were mixed and transplanted into lethally irradiated WT mice (CD45.1/CD90.2; 8–10 weeks old). Each population was tracked by a flow cytometer using congenic markers. Absolute cell numbers of the chimera in the thymus (e) and spleen (f) are shown. (g) Schematic representation of the TCR Vα–Jα–Cα boundary. (h) DP thymocytes were sorted from T-Red and control mice (8–9 weeks old) and isolated from total RNA. RT-PCR was performed using Vα8- and Cα-specific primers followed by southern blotting with Jα- or Cα-specific probes (left). Representative images from three independent experiments are shown. Cβ1 mRNA levels were examined by quantitative PCR (right). Relative expression levels to Hprt are shown. Data represent the mean+s.e.m. (b, n=7–39; c, n=4; d, n=4–6; e,f, n=9; h, n=3). Paired Student’s t-test was used in f. **P<0.01 and ***P<0.001. NS, not significant.