Fig. 5

Bioinformatics combined with in vitro experiments to determine that JUP/AGR2/LYPD3 signaling in melanoma cells can maintain stemness. A Boxplot showing the difference in RNA stemness score between C1 and C2. B UMAP plot of melanoma cells. C, D tSNE plot demonstrating the degree of differentiation of each melanoma cell cluster assessed by CytoTRACE. E Boxplot showing the differentiation score of melanoma cell cluster. F Volcano graph demonstrating the feature genes of each melanoma cell cluster. G UMAP plot showing the co-localization of JUP, AGR2, and LYPD3. H Images of colony formation assay in control, NC, oe-JUP, sh-JUP, and oe-JUP + sh-LYPD3 melanoma cell group. I Images and quantitative analysis of sphere-forming assay in control, NC, oe-JUP, sh-JUP, and oe-JUP + sh-LYPD3 melanoma cell group. J–K Bubble and UMAP plots demonstrating upregulation of glycolysis levels in cluster 1 melanoma cells. L Lollipop plot demonstrating the Spearman correlation between JUP/AGR2/LYPD3 signaling and five key enzymes of glycolysis in TCGA-SKCM. Only SLC2A1 (GLUT1) is consistently and significantly correlated with JUP, AGR2, and LYPD3. (M–N) Verification of GLUT1 expression by Western blotting assay in control, NC, oe-JUP, sh-JUP, and oe-JUP + sh-LYPD3 melanoma cell group. One-way ANOVA was applied. **P < 0.01, ***P < 0.001, ****P < 0.0001