Differential methylation and sex determination. (A) Differentially methylated and differentially expressed genes in the putative sex determination pathway of tongue sole. For each gene presented in the pathway, the methylation (left square) or expression (right square) changes when comparing testes with ovaries are shown by different colors. (B) DNA methylation and transcription of dmrt1 in different developmental stages after hatching. The methylation levels of different stages were estimated using bisulfite-PCR followed by TA-cloning with a pair of primers targeting the first exon, always using at least 10 randomly selected clones for sequencing for each stage. (C) DNA methylation profiles of gsdf in the five gonadal samples. Green vertical lines indicate the methylation level of cytosines. The light gray box indicates the DMR upstream of gsdf. Profiles of other DMGs in the pathway are presented in Supplemental Figure S8.
