Figure 4

T cell subsets in aged Optn^470T were comparable to WT male mice. The gating strategy in mouse spleens is shown for a representative two-year-old mouse (A). Population frequency (%) and absolute cell numbers (No) are shown for one- and two-year-old WT and Optn^470T as follows: frequency of T cells (B), CD8⁺ and CD4⁺ T cells (C), CD8⁺ naïve (T_N), central memory (T_CM) and effector memory (T_EM) (D), and CD4⁺ T_N, T_CM and T_EM (E); numbers of T cells (F), CD8⁺ and CD4⁺ T cells (G), CD8⁺ T_N, T_CM and T_EM (H), and CD4⁺ T_N, T_CM and T_EM (I). Staining for the indicated cytokines upon PMA/ionomycin stimulation is shown as frequencies and MFI in CD8⁺ (J,K) and CD4⁺ (L,M) T cells. The gating strategy for Treg cells in mouse spleens for representative two-year-old mice is shown. Functional Tregs were gated as CD4⁺FOXP3⁺CD25⁺, and non-functional as CD4⁺FOXP3⁺ CD25^- cells (N). The graph shows the MFI of FOXP3 in Optn^470T normalized to one- and two-year-old WT mice in functional Tregs (O). Treg subsets are shown as frequency (P) and numbers (Q). The data from 6–7 one-year- and two-year-old WT and Optn^470T mice are shown as means ± SEM and analysed by two-way ANOVA ((B,C,D): T_N; (E,F,G,H): T_N and T_EM; (I,J,L,P) and (Q), Kruskal–Wallis test ((D): T_CM, T_EM; (H): T_CM) and Student’s t-test (K,M,O): *p < 0.05, **p < 0.01, ****p < 0.0001.