Loss of keratinocyte Dsg1 drives melanoma cell migration. Melanoma cells were cultured for 24 h in conditioned media from keratinocytes transduced with shNT or shDsg1 knockdown vectors and collected for RNA-sequencing. (A and B) GSEA comparing differentially expressed gene sets in RNA Seq run 2 to published signatures of melanoma signaling and differentiation. Normalized enrichment score (NES). (B) Top summary terms from Metascape for overrepresented in genes upregulated by shDsg1 keratinocyte conditioned media. (C) Schematic of trans-well migration assay. Melanoma cells were cultured in conditioned media from shNT expressing keratinocytes or shDsg1 expressing keratinocytes for 24 h. 50,000 cells were then seeded in serum-free media in the upper chamber of a trans-well insert (8- μM pore size). Lower wells contained DMEM supplemented with 5% FBS. (D and E) After 24 h, migrated cells were fixed and stained for visualization. Bars represent normalized migration compared with shNT control conditioned media. Mean ± SEM depicted. n = 3; *P <0.05 Student’s t test.