DNA methylation patterns in XX, XY, and XX + T. Non-CpG methylation increases with age. (a) Global CpG methylation levels are significantly higher at PN60 than PN4 in the BNST/POA but not the striatum. CHG (b) and CHH (c) methylation levels are significantly higher at PN60 than at PN4 in both brain regions. (d, e) The heat maps show correlations between the methylation levels across all samples in the (d) striatum and (e) BNST/POA. The diagonal axis running from the bottom left to top right corner is the line of symmetry where each sample correlates with itself. The brightest yellow shade represents full correlation (=1) whereas the darkest red shade represents the lowest correlation value (0.98; key at top left). Complete clustering is based on the Euclidean distance. (f–i) These heat maps show differentially methylated regions (DMRs) driven by sex and/or testosterone at PN60. The color key shows absolute methylation level from 0% (blue) to 100% (red). The azure trace represents the moving average of the methylation levels in the neighboring windows. (f) Autosomal DMRs in the striatum. (g) Autosomal DMRs in the BNST/POA. (h) X-linked DMRs in the striatum. (i) X-linked DMRs in the BNST/POA.