Dnmt3a enzymes are required for methylation of CAC trinucleotides in the zebrafish brain. (A) Transcript levels of dnmt3aa and dnmt3ab in 4-week-old dnmt3aa/ab CRISPR/Cas9 KO (cKO) zebrafish brains relative to wild type (WT). The data is represented as the mean of technical replicates with error bars showing the standard error (two sample t-test, **P < 0.01). (B) Percentage of reads with no mutation, in-frame mutations, or frameshift mutations, which map to dnmt3aa and dnmt3ab loci in dnmt3aa/ab cKOs. (C) Average mCAC/CAC levels at all CAC trinucleotides and the top methylated CAC motifs in 4-week-old WT and dnmt3aa/ab cKO brain. The data is represented as the average of two WGBS biological replicates (Wilcoxon test, *P < 0.05, **P < 0.01, λ = unmethylated lambda spike in control). (D) mCAC/CAC levels of all gene bodies in 4-week-old WT and dnmt3aa/ab cKO brains. (E) Genome browser snapshot of mCAC/CAC levels in adult brains, 4-week-old WT brains and 4-week-old dnmt3aa/ab cKO brains.
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