FIGURE 5

FIGURE 5

Combined homozygous deletion of nsd1a/b causes derepression of endogenous pericentromeric Sat1 repeats without impacting key molecular markers of heterochromatic silencing. (A) Survival plot comparing wild-type and MZnsd1a/b homozygous deletion mutants over the first 12 months of development (n = ∼60 animals per genotype). (B) Bright field image of representative wild-type and MZnsd1a/b homozygous mutant adult zebrafish at 6 months post fertilization. (C) Expression from pericentromeric Sat1 repeats in wild-type and MZnsd1a/b homozygous mutant larvae at 3 dpf as assessed by qPCR. (D) ChIP-qPCR for IgG, H3K3me2, H3K36me3 and H3K9me3 at Sat1 pericentromeric sequences in wild-type and MZnsd1a/b homozygous mutant larvae at 3 dpf. (E) Southern blot of genomic DNA isolated from wild-type and MZnsd1a/b homozygous mutant larvae at 3 dpf, digested with the methylation sensitive restriction enzyme HpyCH4IV and probed with Sat1 sequence. Equivalent digestion suggests 5 mC levels at Sat1 repeats are unaffected in MZnsd1a/b homozygous mutant larvae. (Wt = Wildtype). Error bars indicate standard deviation.