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Figure 3—figure supplement 1. Analysis of SATB2 expression and chromatin effect in human and zebrafish melanoma.

(A) Western blot analysis in MCR:EGFP, MCR:SATB2, and MCR:SATB1 zebrafish tumors of SATB2 expression and antibody cross reactivity with human SATB1 (in MCR:SATB1) and lack of detection of endogenous zebrafish satb1. The SC-81376 antibody was used for anti-SATB2 ChIP-seq in MCR:SATB2 tumors, since the other anti-SATB2 antibodies shown in Figure 2—figure supplement 3B< did not work in immunoprecipitation. (B) HOMER motif prediction analysis of significant peaks (P<E-9) from anti-SATB2 ChIP-seq enrichment over input in MCR:SATB2 tumors showing the top motif to be AT-rich as expected. (C) HOMER known motifs enrichment analysis on anti-SATB2 ChIP-seq peaks with p<E-9 in MCR:SATB2 (D) Go-term enrichment analysis of genome-wide H3K27Ac peaks enriched in MCR:SATB2 over MCR:EGFP Around TSS Log2FC > 1(GREAT analysis). (E) RNA-seq analysis of satb2 expression in sorted crestin:EGFP+ developing neural crest cells (data source7). (F) Knock-down of satb2 in the transgenic neural crest reporter line Tg(sox10:mCherry) using a validated morpholino for satb2. While satb2 knock down did not affect melanocyte development (0/120), satb2 morphants displayed severe cranio-facial abnormalities as observed by overall reduced sox10 reporter expression, reduced pectoral fins, and head size (109/120) (arrows). (G) Example analysis of the chromatin state of a direct SATB2 target showing the pdgfab locus. ChIP-Seq track in zebrafish MCR:SATB2 for SATB2 (in black). ChIP-seq tracks for H3K27Ac and H3K9me3 in primary tumors MCR:SATB2 (red) and MCR:EGFP controls (green).

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