Figure 3.

Castration increased splenic CD4⁺CD25⁺Foxp3⁺ Tregs by 5 weeks after immunization of wild-type mice. Splenocytes from mice isolated at 2.5 weeks (A) or 5 weeks after immunization (B), and castration or sham treatment were analyzed by flow cytometry. The number of CD4⁺CD25⁺Foxp3⁺ cells was calculated by multiplying the percentage of CD4⁺CD25⁺Foxp3⁺ cells times the total number of isolated splenocytes and represents the average of 3 animals in the group. *, P < 0.01 versus immunization only group. Data are from one representative experiment of 3 independent repeats. Mice were from the experiments shown in Figs. 1 and 2. C, mice were castrated and analyzed 2.5 weeks or 5 weeks later, as described above. Pooled data of 2 independent experiments are shown with 4 to 5 mice per group in each experiment. Cx, castration; Immun, immunization; Sx, sham treatment.