Figure 1. Cytotoxic tumor microenvironments are associated with reduced lymphatic vessel density.

A. Representative immunofluorescence images of LYVE-1⁺ (red) lymphatic vessels from Y1.7 and YR1.7 tumors, nuclei are stained with DAPI (cyan), at day 12 of tumor growth. Scale bar = 100μm. B. Quantification of lymphatic vessel density from (A). C. Gating strategy to identify lymphatic endothelial cells (LEC; CD45⁻CD31⁺gp38⁺). D. LECs as a percentage of total live cells and E. Ki67 as a percentage of total LECs in Y1.7 and YR1.7 tumors. F. Representative IHC images of Lyve1 staining in naïve and Tyr::Cre;Braf ^CA/+;Pten ^fl/lf tumors, G. Quantification of lymphatic vessels in naïve and Tyr::Cre;Braf ^CA/+;Pten ^fl/lf tumors, each dot is a mouse n=4–5, H. Quantification of LECs and BECs as a percentage of total cells per melanoma patient in responders (R) and non-responders (NR) before and on immunotherapy from the single-cell dataset. I. Quantification of LECs in IFNγ⁺CD8 T low and high patient samples from the single-cell dataset, each point is a patient. J. Quantification of LEC signature in IFNγ⁺CD8⁺ T low and high patient samples from the TCGA dataset, each point is a patient. K. Schematic of peritumoral (PT) and intratumoral (IT) lymphatic vessels and T cells. Two-sided, unpaired Student’s t-tests (B-J), Wilcoxon rank test (H). *p<0.05; **p<0.001.