Fig. 3

Validation of sexually dimorphic DNA methylation of the fezf2 gene in Chrysemys picta hatchling gonads by methylation-sensitive restriction enzyme PCR. DNA from three females (1, 2 and 3) and three males (4, 5 and 6) was digested or not with HpaII (HpaII cuts unmethylated DNA). Expected size of the PCR amplicons from PCR primers F1 plus R1, or F1 and R2 are indicated by the arrows (non-specific PCR products were also obtained). L = DNA ladder (1 kb plus). Amplification of the expected fragment in females and not males using the F1 + R2 primers confirms the hypermethylation of fezf2 in females detected using the MeDIP-seq data