Fig. 1: Single-cell analysis of LECs in distant and metastatic LNs from patients with breast cancer.

a Workflow. Distant LNs and paired metastatic LNs from nine individuals were used in this study. Cells were dissociated immediately after surgery, and the presence of tumor cells was verified by staining with pan-cytokeratin. CD45⁺ cells were depleted and PDPN⁺ CD31⁺ LECs were enriched using fluorescence-activated cell sorting. 10x Genomics Chromium scRNA-seq was employed to profile the cells. b Sample information. Patients #2 and #9 had Her2-enriched breast cancer and the others had luminal breast cancer. Paired metastatic and distant axillary LNs were collected from patients #1 to #9. c PDPN and CD31 staining of distant and metastatic LNs. The mean fluorescent intensity (MFI) represents PDPN expression on LECs (distant LN, n = 5 patients; metastatic LN, n = 7 patients) (mean ± SEM, two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file. d UMAP plot of 99,671 LECs from distant and metastatic human LNs, color coded by cluster (left) or metastatic state (right). e Heatmap displaying single-cell expression of the top DEGs in LEC subsets, with selected genes labeled. f Gene expression differentiating the 14 LEC clusters, illustrated in violin plots. AXLN, axillary lymph node.