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. 2023 Jul 22;20:171. doi: 10.1186/s12974-023-02848-0

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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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Protein expression levels in B cells from the spleens of mice after SCI were analyzed by 4D label-free quantitative proteomics. a In the significant difference analysis of the quantitative results, we first selected at least half of the repeated experimental data in the sample group for statistical analysis. Proteins with expression differences of more than twofold and a P ≤ 0.05 are regarded as differentially expressed proteins. The results of the differential protein screening are displayed in the form of a volcanic map, with red spots indicating upregulated proteins (red arrow indicates Tom20), green spots indicating downregulated proteins, and gray indicating proteins with no difference in expression. b GO functions of all differentially expressed proteins were analyzed using Blast2Go software. Blue indicates involvement in biological processes, orange indicates a role in cell composition, red indicates a role in molecular function, and the corresponding box represents the cell function of interest. c Taking the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway as a unit and all qualitative proteins as the background, the significance level of protein enrichment in each pathway was analyzed and calculated by Fisher’s exact test to determine the metabolic and signal transduction pathways that were significantly affected. The red box shows the cellular pathway of interest. d Functional domains of differentially expressed proteins were analyzed using the InterPro database, and the annotated results of differential protein domains were enriched. The red box indicates the domain of interest. e The subcellular localization of differentially expressed proteins was analyzed using WoLF PSORT software. The red box indicates the subcellular location of interest. f In the clustering analysis, a clustering algorithm was used to classify the two dimensions of samples and variables. The red box indicates the Tom20 protein. g Western blotting was used to verify the expression of Tom20 in B cells from the spleens of mice after SCI. The data of Western blotting are expressed as the mean ± SD (n ≥ 3 replicates per group). ** P < 0.01. SCI spinal cord injury, GO gene ontology, KEGG Kyoto Encyclopedia of Genes and Genomes, SD standard deviation