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. 2022 Nov 16;28(4):1571–1584. doi: 10.1038/s41380-022-01862-7

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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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Fig. 2 — a–g ATAC-seq analysis of two-month old control and EtOH-exposed cortical organoids. a Transcription start site (TSS) enrichment plot showing TSS ± 1.0 Kb for each sample. ‘Control 1’, ‘Control 2’, ‘EtOH 1’, and ‘EtOH 2’ labels denote independent batches of organoids made from the WT83 cell line. b Venn diagram depicts peaks in EtOH-exposed and control organoids. c, d Motif enrichment in control (c) and EtOH-exposed (d) organoids. e Plotting enrichment P values identifies prominent regions of altered accessibility. f Tracks plot of select prominent genes. g Reactome analysis predicts that the effects of altered chromatin accessibility concentrate in physiological processes central to neurodevelopment. h, i Mass spectrometry analysis shows histone modifications in astrocytes, cortical organoids, and fetal neurons (hNE) attributable to EtOH-exposure; cortical organoids and astrocytes were generated from the WT83 iPS cell line. Heatmap shows EtOH exposure modifies histone methylation (ME) and acetylation (AC) patterns in diverse neural cell types; scale bar portrays relative abundance of modification across samples: red=high compared to other samples, blue=low compared to other samples (h). A recurrent pattern of modifications is observed between astrocytes, organoids, and fetal neurons; only statistically significant differences in relative modification abundance are shown (i).