Supplementary Figure 6: Immuno-expansion in old Sesn1–/– mice upon vaccination.
From: A sestrin-dependent Erk–Jnk–p38 MAPK activation complex inhibits immunity during aging
(a) Expression of sestrin proteins among CD4+ T cells from young (2 months) or old (20 months) mice assessed by intracellular specific flow-cytometry. Age-dependent expression of Sestrin presented relative to that of young mouse CD4+ T cells, set as 1 (n = 3). (b) Measurement of mouse Sesn1 by both immunoblotting (left) and real-time PCR (right) in splenocytes of indicated genotypes (age: 20 months). Data representative or pooled from 3 separate mice. (c) Spleens from Sesn+/- and Sesn1-/- null mice 5 days after vaccination with FLUAD (1:20 of the human dose). Frequencies of (d) CD4+/ CD8+ T cells (e) myeloid and (f) NK cells in Sesn1+/- and Sesn1-/- mice after vaccination as in (c). (g) Effector/naïve phenotype of CD4+ T cells in control Sesn1-/+ and Sesn1-/- null mice, five days after vaccination with FLUAD. (h) FLUAD-driven IgG isotypic switch in Sesn+/- and Sesn1-/- CD19+ B cells examined by flow-cytometry. (i) Cumulative pre and postvaccination immune-phenotypes of 20-month old control Sesn1+/- and Sesn1-/- null mice. Data (4 old mice per group) presented relative to those of unvaccinated control Sesn1+/-, set as 1. ***p<0.001, ANOVA with Bonferroni post-correction test for comparisons. Error bars depict s.e.m.
