a, Heat map of genes downregulated in zebrafish melanomas that overexpress SETDB1 compared with control (EGFP) melanomas. b, Graphical representation of the rank-ordered gene list derived from a panel of human melanoma short-term cultures stratified based on SETDB1 expression level. GSEA shows that homologs of zebrafish SETDB1-downregulated genes are similarly downregulated in human melanomas as levels of SETDB1 increase (ES = -0.35, NES = -1.43, FDR q-val = 0.045, p = 0.045). Arrows indicate positions of HOX genes. c, SETDB1 and H3K9me3 ChIP-Seq profiles at the HOXA locus in human melanoma cells. The number of sequence reads is shown on the y axis. d, Melanoma-free survival curves of zebrafish expressing SUV39H1 (p = 6.74×10-8 vs. miniCoopR-EGFP, logrank 72) and the methyltransferase-deficient SETDB1(H1224K) (p = 0.24 vs. miniCoopR-SETDB1, p = 8.4×10-5 vs. miniCoopR-EGFP) and SETDB1(C1226A) (p = 0.20 vs. miniCoopR-SETDB1, p = 1.3×10-11 vs. miniCoopR-EGFP) variants. e, In vitro reconstitution of methyltransferase complexes. Sequential purification of GST-tagged SUV39H1, Flag-tagged GLP and HA-tagged G9a proteins was followed by western blotting using antibodies shown on left. f, Histone methylation assays on complexes purified from C2C12 cells.