(A) Total number of male PGCs sorted at E13.5. Error bars, ± SEM; p=0.0274, two-tailed Unpaired t test; n =18 biological replicates. (B) Analysis pipeline to determine which of the de-repressed RNAs at E13.5 had ≥ 20% DNA methylation in wild-type PGCs at E9.5. (C) Heat map showing the methylation status of the 35 de-repressed DEG at E13.5 with promoter methylation at E9.5. (D) Expression levels of de-repressed in common to males and female DCKO PGCs that also had promoter methylation of ≥ 20% at E9.5. FPKM stands for Fragments Per Kilobase of transcript per Million mapped reads. The genes with ≥ 2-fold difference in expression and FDR < 5% were considered differentially expressed. (E) Immunofluorescence of E13.5 male gonads for MVH (red), and STRA8 (green). Included on the right is a positive control for STRA8 staining using an E13.5 DCKO female. See also Figure S3, Table S2, and Table S4.