Figure 2. Impaired antigen-specific T-cell immune response in Hrd1 cKO mice.

WT and Hrd1 cKO mice were immunized with 50 μg KLH-NP in CFA on day 1 followed by a boosted immunization with 50 μg KLH-NP in IFA on day 8. Mice were killed on day 14. (a–c) Total splenocytes were stained with CFSE and cultivated with 100 ng ml⁻¹ of KLH-NP for 5 days. The proliferation (a,b) and apoptosis (c) of CD4⁺ T cells were analysed. The representative images (a,c) and data from five pairs of mice (b) are shown. (d–g) The production of IL-2 in CD4 T cells (d), and TNF-α, IFN-γ and FoxP3⁺ Tregs in CD4 (e) and CD8 (f) cells were analysed. The representative data (d–f) and the mean±s.d. (g) from five pairs of mice are shown. (h) The levels of NP-specific antibodies in the sera of immunized Mann–Whitney test mice were analysed by enzyme-linked immunosorbent assay (ELISA). Error bars represent data from five pairs of mice (mean±s.d.). The Mann–Whitney test was used for the statistical analysis. NS, no significant difference; *P<0.05, **P<0.01 and ***P<0.001.