Fig. 2: CCR7⁺ DCs that migrate to the dLN become phenotypically and transcriptionally distinct from tumour-residing populations.

a–c Flow cytometry of draining lymph nodes (dLN) and contralateral non-draining lymph nodes (ndLN) of MC38-Ova tumours; DCs that originate from photo-flashed tumours (tumour emigrants) carry the Kaede-red fluorescent profile, which enables tracking to LNs. a Representative plots 48 h after tumour photoconversion, b Time-course and quantification of Kaede-red cells among CCR7⁺ DCs, and c composition of Kaede-red DCs in dLNs. d, e UMAP of myeloid cells from scRNA-seq of FACS-sorted CD45⁺ Kaede-red cells from tumour-dLNs (MC38-Ova) and CD45⁺ cells from control LNs. f Gene set enrichment analysis (GSEA) of Kaede-red CCR7⁺ DCs in dLNs versus tumours (isotype control-treated). Signed P-adj indicate log₁₀(Benjamini-Hochberg adjusted P value) with the direction of enrichment. g Selected gene expression in scRNA-seq of tumour CCR7⁺ DCs (isotype control-treated) and Kaede-red CCR7⁺ DC tumour emigrants in the dLN. h Flow cytometry of CD80 and CD86 on CCR7⁺ DCs from MC38-Ova tumours or dLNs 48 h after photoconversion, with representative histograms. i Integration of scRNA-seq of Kaede-red DCs from tumour-dLNs with tumour DCs and label transfer. j Proportion of DC clusters by tissue and Kaede fluorescence from i. Paired two-sided student’s t-test (b, c), two-sided Wilcoxon rank-sum test with Benjamini-Hochberg multiple-testing correction (g), or one-way analysis of variance (ANOVA) and Šidák’s multiple comparisons test (h) were used. Points represent independent mice (b, c, h). Data are shown as means ± s.d. (b, c, h), or box (median; box, 25^th and 75^th percentile; whiskers, 1.5*inter-quartile range) and violin plots (g). The results shown in a–c are from one experiment (24 h/72 h n = 5; 48 h n = 4 animals), representative of three independent experiments; and h from two independent experiments (n = 8 animals).