Figure 6.

Raptor depletion causes reduced switching and affinity maturation in hapten- or tamoxifen-independent systems. (A) Relative concentrations of anti-KLH Ab were measured by ELISA using sera from the eight mice immunized with NP-KLH for analyses of Fig. 5A. Shown are mean (±SEM) ELISA results across serial four-fold dilutions (1:50 to 1:12,800). After screening statistical significance by ANOVA, the null hypothesis was tested pairwise and achieve the indicated P values for IgG1 in samples at the dilutions 1:80, 1:200, and 1:800. (B-E) Cγ1-Cre knock-in mice (Rptor +/+ or f/f, six of each genotype distributed equally in two independent experiments) were immunized as in Fig. 2A. Shown are serological analyses (B) as well as B cell numbers (C), flow staining to measure the frequencies of B cells of GC phenotype (D), or (E) Tfh cells as in Fig. 5D. Shown in (B) are ELISA determinations of circulating anti-NP Ab (all-affinity, left panels; high affinity, panels to right) across serial 10-fold dilutions (1:10 to 1:10⁸), with each point representing mean (±SEM) data for the individual samples (six WT, six mutant). After screening the overall curves by ANOVA, statistical significance was tested pairwise and yielded the indicated P values for samples at the dilutions 1:10 through 1:10⁴. (D, E) Shown are flow data from a representative mouse of each genotype in one experiment, with inset numbers and italicized means (±SEM) as in Fig. 2C.