Extended Data Fig. 1: Decreased migratory cDCs in both human and mouse tdLNs.

a,b, Immunofluorescent staining of cDCs (CD11C and LAMP3), T cells (CD3), or B cells (CD19) in early- or late-stage SLNs from breast cancer (n = 5 vs 6 patients) (a) and colon cancer (n = 5 vs 5 patients) (b) patients. Scale bars, overview, 1 mm; zone a and b, 100 μm. Quantifications on the right panel were CD11C/LAMP3 double positive area out of the total SLN area (%) (upper), or area value of 2 randomly selected areas (500 × 500 μm²) within each sample (lower) (n = 10 vs 12 areas in a; n = 10 areas each in b from corresponding patients). Lower quantification is displayed as area per 10 μm². c,d, Quantifications of tumour volume and tdLN mig-cDCs (CD11c⁺MHC-II^hi) at day 7, 10, and 14 (c, B16F10, n = 4, 4, 6 mice; d, MC38, n = 4 mice per group). e, Quantification of IFNγ and TOX frequencies in CD44⁺PD1⁺CD8⁺ T cells in B16F10 tumour (n = 3 vs 6 mice). f, Quantifications of mig-cDCs, mig-CD103⁺ cDCs, or mig-CD11b⁺ cDCs in tdLNs of MC38 tumour-bearing mice (n = 5 vs 6 mice). g, Flow cytometric gating strategy for DC populations in tdLNs (sample derived from B16F10-OVA-ZsGreen tumour bearing mice). ZsGreen negative control was indicated as grey. Migratory cDC1s or cDC2s were pre-gated on migratory cDCs (CD11c⁺MHC-II^hi). h, Quantifications of migratory XCR1⁺ and SIRPα⁺ cDCs in tdLNs of B16F10-OVA-ZsGreen -bearing mice (n = 6 mice per group). i, Quantifications of migratory CD103⁺ and CD11b⁺ cDCs in non-tdLNs of B16F10-OVA-ZsGreen-bearing mice (n = 6 mice per group). j, Quantifications of total CD8⁺ T cells, CD44⁺CD8⁺ T cells and PD1⁺CD8⁺ T cells in tdLNs from mice bearing early- or late-stage tumours (n = 6 mice per group). Data are merged (a, b) or representative (c-j) of at least two to three independent experiments and present as mean ± s.e.m. Statistical analysis was performed using two-tailed Student’s unpaired t-test.

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