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. 2021 Oct 21;24(11):1542–1554. doi: 10.1038/s41593-021-00923-4

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© The Author(s) 2021

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. 5 — a, Representative images of alkaline comet assay, showing SYBR green stained nuclei of untreated (U) control and C9 hiPSCs or 1 hour (hr) after TOP1i-induced DNA damage (D; Damage) or 1 hr after TOP1i removal (R; Recovery) from 3 independent experiments. b, Confocal microscopy immunofluorescence images displaying DNA damage marker γ-H2AX + foci (green) accumulation in C9 ALI-COs at 150 DIV following 2 Gy gamma irradiation (IR) and quantification. Data expressed as mean ± s.d.; n = 3 ALI-CO sections (> 120 cells for each); Two-tailed unpaired t-test. c and d, Representative confocal immunofluorescence images of γ-H2AX⁺ foci in CTIP2⁺ DLN (c) and SOX9⁺ astroglial nuclei (d) in C9-L2 vs ISO-L2 ALI-COs treated with TOP1i alone ± ATM kinase inhibitor, AZD0156 (ATMi). Quantification of γ-H2AX⁺ foci. Data expressed as individual foci/cell, mean ± s.d.; n = 95-152 DLNs (c) and n = 99-193 astroglia (d) from 3 independent ALI-COs per group. Kruskal-Wallis test with Dunn’s multiple comparisons test. Scale bar=5 µm for all images. See Supplementary Table 3 for detailed statistics.