Fig 2: Human DC Heterogeneity, Related to Figure 7(A). Violin plots showing expression distribution of mouse DC subset marker genes across human peripheral blood DC and monocyte clusters identified in Villani et al. (2017).(B). Representative flow cytometric analysis of mouse peripheral blood cDC2s showing absence of T-bet (RFP)+ cDC2s.(C). Gating strategy for FACS-isolation of human spleen DCs for scRNA-seq. DCs were defined as live, LIN(CD3,CD56,CD19)-CD14–CD11C+HLA-DR+.(D). Representative flow cytometry analysis of human spleen cDC2s gated as Lin(CD3,CD56,CD19)–CD14–CD11c+HLA-DR+CD123–XCR1–CLEC4A+ cells. Left panel: cell surface expression of CD1c and CLEC10A by cDC2s. Right panel: overlay of CLEC10A+ and CLEC10A– cDC2s distinguished by differential expression of CLEC4A and FceR1a. Summary bar graphs show frequency of CD1C+CLEC10A+ and CD1C+CLEC10A– cDC2s as a percentage of cDC2s (n = 4 individuals).(E). t-SNE embedding of 9,315 FACS-isolated CD45+ immune cells from two melanoma tumors. Colors indicate unsupervised clustering by Phenograph (left panel) or classification based on expression of canonical markers and correlations with bulk RNA-seq data (right panel). Each dot represents an individual cell.(F). Pearson correlations between cluster centroids in (F) and bulk RNA-seq data from purified immune populations (Jeffrey et al., 2006, Novershtern et al., 2011)(G). t-SNE map of 2,122 myeloid cells identified in (F). Colors indicate patient sample (left) or unsupervised clustering by Phenograph (right panel). Each dot represents an individual cell.(H). Heatmap of normalized, log transformed and MAGIC imputed expression of top 20 differentially expressed genes, defined by the highest earth mover’s distance (EMD), per Phenograph cluster in E. The colored bar at the top of the heatmap shows assignment of cells to clusters labeled in F, right panel.(I). t-SNE map of human melanoma myeloid cells (H) colored by imputed expression of labeled genes.