Fig. 4

Transcription is required for DNAme targeting at the Zac1 locus. a Visualization of the Zac1 transcripts in somatic tissues (top) and in oocytes (bottom), as well as the DNAme landscape at this locus in FGOs. Deletion of Zac1o promoter is indicated by del. above the Cufflinks annotation, and below the DNAme profile are indicated the regions (IN1, IN2, IN3, igDMR) that are subsequently assessed for DNAme in (b, c). b DNAme status of Zac1 igDMR and Zac1o/Zac1oAS intragenic regions in Zac1o+/+ and Zac1o−/− FGOs. DNAme was assessed by bisulfite sequencing (BS-PCR) and each line represents an individual unique clone, with open circles representing unmethylated CpGs and closed circles methylated CpGs. c DNAme (BS-PCR) status of the Zac1 igDMR and Zac1o IN2 intragenic region in Zac1o+/+ and Zac1o+/− neonatal (postnatal day 2 (P2)) brain. d Sequence traces (left) of RT-PCR products from neonatal brain from Castaneus crosses to Zac1o+/+ and Zac1o−/−; the asterisk indicates the T/C single-nucleotide polymorphism. Zac1o and Zac1 expression assessed by quantitative RT-PCR (right) in Zac1o+/+ and Zac1o+/− neonatal brain (***p < 0.001, **p < 0.01, Student’s t-test). e ChIP-quantitative PCR quantification of H3K4me2 and H3K36me3 enrichment in growing oocytes (15 dpp) at Zac1 igDMR, Zac1o intragenic regions and Zac1o intergenic regions (ND non-determined, *p < 0.05, **p < 0.01 Student’s t-test)