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. 2024 Nov 15;44(1):1–29. doi: 10.1038/s44318-024-00298-9

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) qRT-PCR-based gene expression measurements (n = 3) for APOBEC3A (P = 0.0002), KRT10 (P = 0.0002), IVL (P < 0.0001), APOBEC3B (P = 0.0001), MKI67 (P < 0.0001) and MCM7 (P = 0.0033) in proliferating NIKS (Async) or following 48 h of growth factor deprivation (Starved). (B) Representative cell cycle profiles of Async and starved NIKS measured by PI staining and flow cytometry. (C) qRT-PCR-based gene expression measurements (n = 3) for APOBEC3A (P = 0.0036), KRT10 (P = 0.0004), IVL (P = 0.0007), APOBEC3B (P = 0.9445), MKI67 (P = 0.0032) and MCM7 (P = 0.0001) following 24 h of vehicle control (DMSO) or 100 nM afatinib treatment (EGFRi). (D) Representative cell cycle profiles of DMSO and afatinib-treated NIKS measured by PI staining and flow cytometry. (E) qRT-PCR-based gene expression measurements (n = 3) for APOBEC3A (adjusted P values versus day 3 measurement: day 5 = 0.9960; day 7 = 0.0009; day 9 <0.0001), KRT10 (adjusted P values versus day 3 measurement: day 5 = 0.9518; day 7 = 0.0005; day 9 <0.0001), IVL (adjusted P values versus day 3 measurement: day 5 = 0.9983; day 7 = 0.0035; day 9 <0.0001), APOBEC3B (adjusted P values versus day 3 measurement: day 5 <0.0001; day 7 <0.0001; day 9 <0.0001), MKI67 (adjusted P values versus day 3 measurement: day 5 = 0.0001; day 7 <0.0001; day 9 <0.0001) and MCM7 (adjusted P values versus day 3 measurement: day 5 <0.0001; day 7 <0.0001; day 9 <0.0001) in NIKS collected 3, 5, 7, or 9 days after plating. (F) Representative cell cycle profiles of NIKS collected 3, 5, 7, or 9 days after plating were measured by PI staining and flow cytometry. (G) qRT-PCR measurements of APOBEC3A expression in primary human epidermal keratinocytes (NHEK) growing in FC medium and treated for 24 h with vehicle control (DMSO) or 100 nM afatinib (EGFRi, P < 0.0001) or following growth factor deprivation for 48 h (Starved, P = 0.0003). (H) Percentage of DDOST transcripts that were C > U edited at c558 in asynchronous growing NIKS (Async) and following 48 h of growth factor withdrawal (starved) measured by digital PCR assay (n = 3; P < 0.0001). Error bars = SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Pairwise comparisons were performed using unpaired two-tailed t-tests in (A, C, H). Comparisons of mRNA levels on days 5, 7 and 9 to day 3 in (E), and of mRNA levels in starved and afatinib-treated cells to DMSO-treated cells in (G) were performed using one-way ANOVA with Dunnett’s multiple comparisons test. Source data are available online for this figure.