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. 2023 Aug 18;23:775. doi: 10.1186/s12885-023-11260-z

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© The Author(s) 2023

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Inhibition of PRMT5 increased markers of DNA damage in replicating cancer cell lines. A, The change of the percentage of GSK3326595 treated cells positive for geminin, a marker of replication, and γH2AX, a marker of DNA damage, from DMSO or etoposide treated cells measured by immunofluorescence. Cells with a geminin MFI > 750 were considered geminin positive and cells with an MFI > 2000 or > 15 foci per nuclei were considered γH2AX positive. The change in the percentage of cells positive for both geminin and γH2AX was plotted (n = 2 per cell line with 2 technical replicates per cell line). B, Representative immunofluorescent images of CaOV3 cells treated with DMSO or GSK3326595 for 6 days. Cells stained with anti-γH2AX, anti-geminin, and nuclear stain Hoechst. Immunofluorescent images taken using a 40X objective. Scale bar represents 50 μm. C, The change in the percentage of GSK3326595-treated cells positive for geminin and RAD51, a marker of HR mediated DNA repair, from DMSO or etoposide-treated cells measured by immunofluorescence. Cells with > 5 foci per nuclei were considered RAD51 positive (n = 2 biological replicates with 2 technical replicates per cell line). D, The change in the percentage of GSK3326595 treated cells positive for geminin and 53BP1, a marker of NHEJ mediated DNA repair, from DMSO or etoposide treated cells measured by immunofluorescence. Cells with > 5 foci per nuclei were considered 53BP1 positive (n = 2 per cell line). All error bars represent mean ± SEM.