Fig. 6

Characterization of CD73 positive pro-metastatic NK cells in uveal melanoma A Frequencies of CD73 positive NK cells after 4 h co-culture with either parental OCM-3 or 4-1BBL-KO OCM-3. Wilcoxon signed-rank test was used to test for significance (n = 5). B Proportions of CD73 positive versus negative NK cells expressing NKp46, PD-L1, IL-10, NKG2D and LAG-3. Mann–Whitney test was used to determine the significance (n = 4). C Representative images of red fluorescent labelled MEL270 localization within zebrafish larvae. White scale bar denotes 100μm. D Number of tumor foci quantified within every zebrafish larva. Student t-test was used to determine significance comparing xenografts injected with either CD73 positive or negative NK cells (n = 5). E) tSNE projection of NK cells from primary uveal melanoma tumors (GSE 139829, n = 8) colored based on Louvain clusters (left) and the normalized gene expression of CRTAM, CD69 and PRF1. F Trajectory analysis using monocle 2 algorithm which reveals transitional subclusters (left) in relation to Louvain clusters (Right). G Hierarchical heatmap clustering of differentially expressed genes comparing the 3 louvain clusters. Genes in red font are upregulated in cluster 3 but not related to lipid metabolism. H) tSNE projection of NK cells and their normalized expression of ENTPD1 (top) and TGFB1 (Bottom)