Both proximal and distal IC-LN–derived CD8 T cells contribute to the generation of the memory recall. (A) Experimental design described in Fig. 4 A. Representative flow plot of CFSE fluorescence and Ki67 expression by the retransferred proximal and distal OT-I T cells, as well as by the endogenous polyclonal CFSE-labeled donor and unlabeled recipient CD45.2⁺ CD8 T cells isolated from the spleen (n = 3–4, two experiments). (B and C) Representative plots and quantification of (B) cleaved caspase 3 and (C) BCL2 expression. (D–G) Experimental design described in Fig. 4 D. (D) Representative flow plots of proximal and distal IC-LN–derived OT-I T cells isolated from the spleen following recall. (E) Cellular ratios of distal to proximal IC-LN–derived OT-I T cells. Dotted line represents the original 1:1 ratio at retransfer (n = 5–7, four experiments). (F) OT-I T cell numbers from individual memory recall experiments in naïve and 2′ immunized recipient mice. (G) Quantification of the expression patterns for the indicated cell markers by retransferred OT-I T cells following recall in naïve recipient mice (n = 5, three experiments). Data from multiple pooled experiments are denoted by different symbols within the same group. Graphs show mean ± SD and were analyzed using paired Student’s t test for comparison within the same mice or unpaired for comparisons between mice. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; P > 0.05 not significant (ns).