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. 2020 Mar 26;7:104. doi: 10.1038/s41597-020-0449-z

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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. 1 — Sample preparation and data analysis workflows used in the generation of the spectral library. L929 cell lysates were first fractionated with size-exclusion chromatography and digested with trypsin. The resulting peptides were fractionated with HILIC (Hydrophilic Interaction Liquid Chromatography). The tissue samples were digested with trypsin and the peptides were fractionated with high-pH chromatography. The peptide fractions were dissolved in 0.1% formic acid containing iRT peptides, which were analyzed using shotgun MS. The DDA files were searched with X!Tandem and Comet, and results were combined with iProphet. The combined results were filtered with 1% protein FDR and made a consensus spectral library with Spectrast software.