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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. 4 — a, b Western blot and quantification of astrocytic and synaptic protein markers in cortical organoids during development (two-way ANOVA for each protein; *P < 0.05, **P < 0.01, ***P < 0.001; n = 3 replicates (~5–10 organoids)/condition from independent organoid batches from the WT83 iPS cell line). c Principal components analysis of protein digital spatial profiling distinguishes distinct clusters between proliferative and non-proliferative regions and between EtOH-exposed and control samples (n~10 control and EtOH-exposed organoids) (d) Heatmap showing variation in protein abundance in organoid samples between distinct spatial and EtOH-exposure clusters. e Volcano plot of proliferative (rosette) regions from control and EtOH-exposed organoids highlights the differences in expression of various proteins. Green bar represents a P value of 0.05.