Extended Data Fig. 4: IL-4 present during CD8⁺ T cell activation impairs CD103 expression and CD8⁺ T_RM cell formation/persistence in vivo.

(a) Numbers of CD45.1⁺CD44^hiCD8⁺ and CD45.2⁺CD44^hiCD8⁺ T cells that were in vitro-activated with IL-4 (IL4^Act OT-I) or without (noIL4^Act OT-I), adoptively co-transferred intravenously into congenic CD45.1⁺CD45.2⁺ C57Bl/6 mice that were immediately painted on the flank with DNFB, and recovered from the spleen, bLN and flank skin at day 5 post-transfer. *p = 0.0234, **p = 0.0039, two-sided Wilcoxon test. (b) Numbers of IL4^Act OT-I and noIL4^Act OT-I CD8⁺ T cells recovered from spleen, bLN and flank skin as in a at day 30 post-transfer. *p = 0.0039, two-sided Wilcoxon test. (a, b) Data are pooled from three independent experiments on day 5 (n = 9) and on day 30 (n = 9). (c) Numbers of IL4^Act OT-I and noIL4^Act OT-I CD8⁺ T cells T cells that were adoptively co-transferred intradermally into congenic CD45.1⁺CD45.2⁺ C57Bl/6 and recovered from the bLN and flank skin at day 2 post-transfer. (d) Numbers of IL4^Act OT-I and noIL4^Act OT-I CD8⁺ T cells recovered from spleen, bLN and flank skin as in c at day 5 post-transfer. (e) Numbers of IL4^Act OT-I and noIL4^Act OT-I CD8⁺ T cells recovered from spleen, bLN and flank skin as in c at day 30 post-transfer. *p = 0.0391; **p = 0.0156, two-tailed Wilcoxon test. (f) Percent CD49a⁺, CD69⁺ and CD127⁺ cells of noIL4^Act or Il4^Act OT-I CD8⁺ T cells isolated from the flank skin as in c on day 30 post-transfer. (g) gMFI of CD49a, CD69 and CD127 expression in noIL4^Act and Il4^Act OT-I CD8⁺ T cells isolated from the flank skin as in c on day 30 post-transfer. (c-f) Data are pooled from three independent experiments on day 2 (n = 6), day 5 (n = 7), and day 30 (n = 8).

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