Supplemental Figures 1-20 from A Quantitative System for Studying Metastasis Using Transparent Zebrafish

Supplemental Figures 1-20. Supplemental Figure 1: Metastatic behavior of primary melanoma transplants. Supplemental Figure 2: Demonstration of cDNA overexpression in the ZMEL1 line. Supplemental Figure 3: Demonstration of CRISPR/Cas9-mediated mutation in the ZMEL1 line. Supplemental Figure 4: SURVEYOR assay to assess mutation rates in ZMEL1 cells nucleofected with CRISPR/Cas9 constructs. Supplemental Figure 5: Gene Set Enrichment Analysis (GSEA) for the ZMEL1 transcriptional signature compared to the NCI60 human cancer cell lines (dataset NCI60GSE5846). Supplemental Figure 6: Heatmap plot of the ZMEL1 transcriptional signature compared to the NCI60 human cancer cell lines (dataset NCI60GSE5846). Supplemental Figure 7: Gene Set Enrichment Analysis (GSEA) for the ZMEL1 compared to transgenic mitf-BRAF;p53 tumors. Supplemental Figure 8: Heatmap plot of the ZMEL1 transcriptional signature compared to transgenic mitf-BRAF;p53 tumors. Supplemental Figure 9: Ingenuity Pathway Analysis of the ZMEL1 expression signature showing canonical pathways. Supplemental Figure 10: Ingenuity Pathway Analysis of the ZMEL1 expression signature showing core enriched networks. Supplemental Figure 11: Heatmap of metastatic spread over 14 days when cells are transplanted dorsally. Supplemental Figure 12: Histologic assessment of fish transplanted with ZMEL1-GFP cells Supplemental Figure 13: Histology of the kidney marrow from a stable transgenic mitfa-BRAFV600E;p53-/- melanoma bearing fish. Supplemental Figure 14: Derivation of the µ score for quantifying metastatic burden Supplemental Figure 15: Calculation of the metastasis initiating cell (MIC) frequency Supplemental Figure 16: Schematic overview of bright field image processing. Supplemental Figure 17: Determining pixel intensities in the transformed images. Supplemental Figure 18: Methods for segmentation of GFP images. Supplemental Figure 19: Methods for clustering of close objects. Supplemental Figure 20: Methods for separating merged tumors/metastases.