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. 2022 May 4;606(7912):153–159. doi: 10.1038/s41586-022-04686-1

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 11 — a, Hierarchical clustering of log2FC values for P4 sex DEGs detected in Esr1+ inhibitory neuron clusters (see also Fig. 3e). Sex DEGs with non-significant differential expression colored in white. b, Neonatal E2 (NE) vs. neonatal vehicle (NV) female nuclear RNA-seq on P4 BNST Esr1+ cells; grey, red dots (DESeq2, padj < 0.1). c, (top) Overlap between NE-induced genes and P4 multiome male-biased genes. (bottom) Overlap between NE-downregulated genes and female-biased genes. d, (left) Pearson’s correlation coefficient values for non-differential (grey) and NE-regulated (gold) ATAC peaks that correlate with P4 sex DEG expression. Boxplot center=median, box boundaries=1st and 3rd quartile, whiskers=1.5*IQR from boundaries. n = 5169 non-differential, 244 NE-regulated. p-value from two-sided, Wilcoxon rank-sum test. (right) Distance between non-differential (grey) and NE-regulated (gold) ATAC peaks to P4 sex DEG transcription start sites (TSS). p-value from Kolmogorov-Smirnov test. e, Example P4 sex-biased genes that are also NE-regulated, Htr4 (top) and Csgalnact1 (bottom). (left) n = 3, (right) n = 887 i1:Nfix female cells, 676 i1:Nfix male cells, 404 i3:Esr2 female cells, 550 i3:Esr2 male cells. f, Tracks for NE-open ATAC peaks that correlate with NE-regulated, sex-biased targets, Htr4 and Csgalnact1. g, Different NE-open ATAC peaks across i1:Nfix, i3:Esr2, and i4:Bnc2 neurons correlated with a common male-biased target, Arid1b. h, Heatmaps indicating (left) RNA log2FC of P14 sex DEGs and (right) Pearson’s correlation coefficient of NE-open (red) and -close (blue) ATAC peaks linked to sex DEGs within each cluster. Non-significant genes and correlation values colored in white.