Fig. 1. CHD3 and CHD4 are required for normal melanoma cell proliferation.

a, b 501Mel cells were transfected with the indicated siRNAs and CHD3 and CHD4 expression evaluated by RT-qPCR or by immunoblot along with that of MITF and SOX10. c The indicated cell lines were transfected with siRNA and after reseeding the number of colonies counted after 10 days. d The indicated cell lines were transfected with siRNAs and cell proliferation evaluated by cell trace violet assay. e The indicated cell lines were transfected with siRNA and apoptosis detected by FACs after labelling with Annexin-V. Silencing of MITF known to induce cell cycle arrest and senescence was included as a control. f Volcano plot showing changes in gene expression following CHD4 silencing. Genes up or down-regulated based on Log2 fold-change >1/<-1 with an adjusted p-value <0.05 were identified. g Ontology analyses of CHD4 regulated genes. Shown are the enrichment scores for GSEA, as well as David functional enrichment and KEGG pathway categories. h–j Verification of deregulated expression of selected genes following siCHD4 in independent RNA samples from 501Mel, MM117 and Sk-Mel-28 cells. In all experiments n = 3 biological replicates and unpaired t-test with two tailed p-value analyses and confidence interval 95% were performed by Prism 5. p-Values: *p < 0.05; **p < 0.01; ***p < 0.001. Data are mean ± SEM.