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. 2023 Oct 19;14:6610. doi: 10.1038/s41467-023-42365-5

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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/.

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Fig. 3 — a Gene Ontology annotation of proteins expressed at different levels before and after nsEP treatment. b Gene Ontology enrichment of cellular components. c STRING-based protein–protein interaction (PPI) network analysis of identified proteins with higher expression confidence of 0.7; proteins of interest are displayed as a single diamond-shaped node, and differential proteins that interacted with nodes were analysed by Kyoto Encyclopaedia of Genes and Genomes pathway. d Proteomics proteins volcano plot analysis for sEVs derived from nsEP-treated MEFs. Red triangles and blue diamonds represent proteins associated with the induction of sEVs, among which red indicates proteins that were more highly expressed than others. e Heatmap of the top 95 proteins differentially expressed after nsEP treatment. NA indicates UPF0600 protein C5orf51 homologue (a protein whose gene name is unknown). Proteins associated with sEV secretion are highlighted. f Representative western blots of MLKL identified in the indicated treatment conditions. Repeated three times independently with similar results obtained. g Numbers of sEVs per cell produced by MLKL knockdown or MLKL-overexpressing MEFs. Equal amounts of protein were used for the proteomic analysis, and data were from three independent biological replicates. Data in g represent means ± SD, n = 3 biologically independent samples; statistical significance was analysed with unpaired two-tailed Student’s t-tests. Source data are provided as a Source Date file.