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. 2021 Nov 24;12:6813. doi: 10.1038/s41467-021-27057-2

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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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Fig. 4 — CLB-BAR NB cells were synchronised by thymidine block and treated with either DMSO (Ctrl) or 50 nM BAY 1895344. a Immunoblotting with anti pATR, ATR, pFOXM1, FOXM1, pCHK1, CHK1 and tubulin antibodies after treatment for 6–12 h as indicated. NS, non-synchronised; S, synchronised. b–g Phosphoproteomic analysis. b Differential phosphorylation (DP) upon 6 h BAY 1895344 treatment was determined by comparing the phosphoproteomic response with control (DMSO) conditions. See Supplementary Dataset 1 for the detailed results. c Volcano plot showing DP response. DP sites indicated in blue. ATR and ATM sites indicated and labelled in red. Other sites that are discussed in the main text in black. S/TQ sites indicated by red borders. d Linear motif analysis. Sequence logo plots showing position-specific enrichment ±5 amino acids centred around the phosphorylation site for hypo- and hyperphosphorylated sites as indicated. Pie charts showing the proportion of S/TQ motifs in hypophosphorylated, hyperphosphorylated and non-DP sites. p Value calculated using Fisher’s exact test, comparing S/TQ sites in DP versus non-DP sites. e Heatmap comparing z-score normalised phosphorylation signals between control (DMSO) and BAY 1895344 conditions for hypophosphorylated sites that are discussed in the main text. S/TQ sites labelled in red. f GSEA on 31 proteins with hypophosphorylated sites and 417 proteins with hyperphosphorylated sites. Bar plot showing a selection of Canonical pathways that were enriched at 5% FDR (q < 0.05). See Supplementary Dataset 2 for the detailed results. g STRING network analysis. Subnetworks showing all 1st-degree interacting proteins with hypophosphorylated sites. Log2 (fold change) values labelled by heatscale (key indicated on bottom right). S/TQ sites indicated by red borders. h Schematic highlighting key targets of ATR activity in NB cells. Source data are provided as a Source Data file.