Fig. 1. Postnatal epigenetic maturation in ARH neurons is sex specific.

(A) Illustration of ARH microdissection approach (left) and representative scatterplot from FANS (right). (B) UMAP clustering of autosomal RNA-seq and WGBS data on ARH neurons and glia from both ages and sexes. (C) Miami plot showing the significance of P35 to P12 differential methylation, at the CpG level, in male and female neurons. Points plotted above/below the origin represent CpGs that gained/lost methylation from P35 to P12, respectively. Y axis represents raw P values. Genome-wide significance threshold (horizontal dotted lines) = P < 2.45 × 10⁻⁹. (D) CpG-level plots of example regions from (C) showing clusters of male-specific (i), common (ii), and female-specific (iii) differentially methylated CpGs. (E) Venn diagram illustrating the overlap of mDMRs identified in male and female ARH neurons. Histograms (right) show the magnitudes and directions of these maturational methylation changes. Median differential methylation for P35 > P12 mDMRs ranged from 12.3 to 15.3%, and 13.8 to 17.5% in P35 < P12 mDMRs. (F) Heatmaps showing P35 to P12 differential methylation in 6-kb genomic regions centered on neuron mDMRs; most do not show methylation changes in glia.