Fig. 5
Global molecular signatures in melanoma derived from a defined population of McSCs. a Schematic representation of the strategy for results in Figs. 5 and 6. McSC isolation, transplantation and bulk and single cell RNA-seq comparing McSCs to their derived tumor cells. b Images of mice injected with McSCs from Tyr-CreER:Braf:Pten:Tomato (left) and c-Kit-CreER:Braf;Pten:Tomato (right) mice at 47 days following transplantation. Arrows point to the injection site. Scale bar: 1 cm. c Images of mice injected with wildtype McSCs at 47 days following transplantation. Arrows point to the injection site. Scale bar: 1 cm. d H&E and immunofluorescence for Tomato, S100b and Sox10 in tumors derived from McSCs of Tyr-CreER:Braf:Pten:Tomato (Top panel) and c-Kit-CreER:Braf;Pten:Tomato (Bottom panel) mice at 47 days following transplantation. Scale bar, 50 µm. e RNA-Seq heat map of differentially expressed genes (DEGs) shown comparison of Tyr-CreER:Braf:Pten:Tomato McSCs derived tumors to non-transformed control McSCs. Shown are DEGs with FDR < 0.02 and changes greater than 4 fold between Tyr-CreER:Braf:Pten:Tomato McSCs derived tumors and quiescent McSCs from control Tyr-CreER; R26R-Tomato mice. Red indicates high expression, while blue indicates low expression. Note that because Tyr and c-Kit promoters target the exact same population of McSCs, we chose to do RNA-seq with McSCs isolated from only one of them. f GSEA plots comparing the correlation of a rank-ordered list of McSC-derived tumor enriched genes to the KEGG-extracellular matrix interaction pathway (left), neural crest (middle) and neuronal gene (right) signatures. NES: normalized enrichment score. g Bar graph showing log2 fold changes of indicated markers in melanoma cells compared to normal McSCs identified in RNA-seq. h Immunohistochemical validation of specific RNA-seq targets in quiescent McSCs in telogen hair follicles of control mice (left) and Tyr-CreER:Braf:Pten:Tomato McSC-derived tumors (right). Inset shows separate channel images with higher magnification of boxed area. Scale bar, 25 µm; inset scale bar, 10 µm. Source data are provided as a Source Data file
