Figure 2. Zebrafish scRNA-sequencing shows upregulation of EMT-TFs in tumor-associated keratinocytes.

(A) Schematic of scRNA-sequencing experiment. Embryo injection with miniCoopR:eGFP plasmid and tol2 mRNA in Casper Triples (mitfa−/−;mitfa:BRAFV600E;tp53−/−;mpv17−/−) results in melanocyte rescue and subsequent melanoma formation. Melanoma was dissected and dissociated to single cell suspension for FACS isolation of eGFP+ melanoma cells and non-fluorescent TME cells for single cell RNA-sequencing.

(B) Dimensionality reduction and subsequent analysis with zebrafish keratinocyte gene module scoring highlights two keratinocyte clusters.

(C) Top 6 GSEA Hallmark pathways enriched in comparison between keratinocyte clusters.

(D) Hallmark EMT pathway enrichment in keratinocyte clusters.

(E) Expression of EMT-transcription factors Snail (snai1a), Slug (snai2), and Twist1 (twist1a) in TAK vs. NKC, Melanoma, and Other TME cells. ns is no significance, * is p ≤ 0.05, **** is p ≤ 0.0001.