Extended Data Fig. 5: Dynamic and spatial characterization of OT-II CD45.1+ T cells in murine TME niche and cLN.
(a) Representative FACS plot showing gating strategy for isolation of adoptively transferred CD45.1+ OT-II cells and bystander polyclonal TCRβ+ T cells from tdLN. (b) Quantification of the percentage of CD45.1+ OT-II T cell out of the entire TCRβ+ population in matched cLN and tdLN tissues. Two-tailed paired Mann–Whitney test; n = 8 mice from two independent experiments (c) Gene module analysis. Shown are seven correlated gene modules derived from CD45.1+TCRβ+ OT-II T cells isolated from cLN, dLN and TME, 10- and 17 days post tumor cell injection. Left – pairwise Pearson correlation. Right – normalized pooled expression across different conditions. (d) Pooled expression of the seven gene modules from (c) across OT-II T cells derived from all conditions. Dots represent single cells and dot colors indicate time points. Representative genes from each module are depicted. (e) Fraction of proliferating cells in each quartile of the mTht activation signature as in Fig. 5b in dLN OT-II T cells. A cell was determined proliferating if it exhibited above-threshold expression of the cell-cycle module in (d). (f) Representative confocal microscopy images of cLN sections extracted 10- and 17 days following tumor cell injection, stained for CD45.1+ (OT-II) T cells, PD-1 and CD11c proteins. Scale bar=30μm; asterisks indicate OT-II mTht; images are representative of two independent experiments.
