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. 2020 Jul 16;11(7):538. doi: 10.1038/s41419-020-2727-2

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© The Author(s) 2020

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 U2OS cells were treated with 1 μM ISRIB and at the same time incubated with CldU (30 min). Cells were labeled with IdU (60 min) in the presence of ISRIB and 4 μM Thap and then harvested for DNA fiber assay analysis. b Cells were pre-treated with 1 μM ISRIB for 30 min and then with 10 μM BEPP or 25 μM Sephin in the presence of ISRIB for 2.5 h. To label newly synthesized DNA, cells were incubated with CldU and IdU during the last 1.5 h as shown, and then harvested for analysis. c–e Representative DNA tracks as labeled in red (CldU) and green (IdU) of cells treated with ISRIB/Thap (c), ISRIB/Sephin (d), or ISRIB/BEPP (e). f–h Fork progression of IdU label of cells treated with ISRIB/Thap (f), ISRIB/Sephin (g) or ISRIB/BEPP (h) represented as 5–95 percentile box plots. Plots shown are a representative of two or three independent experiments. See Supplementary Fig. S2A–E.