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. 2024 Sep 25;634(8034):712–720. doi: 10.1038/s41586-024-07962-4

Extended Data Fig. 2. Fc–IL-4 enriches CD8+ TTE cells in an antigen-dependent manner.

Extended Data Fig. 2

a-c, Experimental setting was similar to that described in Fig. 1a (n = 5 animals). Shown are the representative flow cytometry plots (a) and the frequencies (b) of CD8+ TTE cells (TCF-1-TIM-3+) among PMEL or endogenous CD8+ TILs, and frequencies of Granzyme B+IFNγ+ among different subpopulations of PMEL or endogenous CD8+ TILs (c). d-g, CD45.2+ C57BL/6 mice were sublethally lymphodepleted on day -4 and received adoptive co-transfer of CD45.1+ naive OT1 T cells (2 × 106, i.v.) and CD90.1+ naive PMEL T cells (2 × 106, i.v.) on day -3. The mice were then inoculated with B16F10 tumour cells on day 0. On day 7, the mice were treated with ACT of activated CD90.2+ PMEL T cells (5 × 106, i.v.) followed by administration of Fc–IL-4 (20 µg, p.t.) or PBS every other day for 4 doses. On day 15, mice were euthanized and tumour tissues were collected for flow cytometry analysis (n = 6 animals). Shown are the experimental timeline (d), representative flow cytometry plots (e), frequencies of transferred CD45.1+ OT1 and CD90.1+ PMEL T cells among total CD8+ TILs (f), and frequencies of PD-1+TIM-3+ subpopulation among transferred CD45.1+ OT1 or CD90.1+ PMEL T cells (g). h-j, Experimental setting was similar to that described in Fig. 1a (n = 5 animals). Shown are representative flow cytometry plots (h) and frequencies of IFNγ+ or IL-4+ among PMEL and endogenous CD8+ T cells (i), and frequencies of IFNγ+ or IL-4+ among endogenous CD4+ T cells (j). All data represent mean ± s.e.m. and are analysed by two-way ANOVA and Sidak multiple comparisons test (b, and c), or two-sided unpaired Student’s t-test (f-j). Schematic in d created using BioRender (https://Biorender.com).

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