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. 2021 Jan 11;12:254. doi: 10.1038/s41467-020-20509-1

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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. 3 — a Workflow for ubiquitinome analysis in tumor necrosis factor (TNF) signaling. b Identified diGly sites (±SD) for TNF treated (100 ng/ml for 10 min) and control U2OS cells in data-independent acquisition (DIA, blue) and data-dependent acquisition (DDA, red) experiments (n = 6, three workflow replicates measured in analytical duplicates). c Volcano plot of significantly regulated diGly sites at 5% false discovery rate (FDR) (FDR controlled, two-sided t-test, randomizations = 250, s0 = 0.1) (lower line) and 1% (upper line) for DIA (blue) and DDA (red) and overlaps of significantly upregulated diGly sites for 1 and 5% FDR cutoffs (t-test, s0 = 0.1). d Overrepresentation analysis of gene ontology biological process (GOBP) terms filtered for 5% corrected FDR (Fisher’s Exact test). e Cytoscape network of proteins with significantly upregulated diGly sites in DIA that are associated with NFκB signaling (GO 0043122; GO 0051092; 5% FDR). Upregulated diGly sites also captured by DDA are marked in red (5% FDR). Source data are provided as a Source data file.