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. 2017 Oct 17;7:13342. doi: 10.1038/s41598-017-13592-w

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© The Author(s) 2017

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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Impact of β5i/LMP7-deficiency on proteasome composition and proteolytic activity in cells and tissues of LDLR^−/− mice. (A) Western blot analysis of standard proteasome (β1, β2, and β5) and immunoproteasome (β1i, β2i, and β5i) subunit expression in BMDM of LDLR^−/−LMP7^−/− and LDLR^−/− at baseline and after treatment with IFN-γ (100 U/ml) over 8 hours (left panel). Right panel shows Western blot analyses of pooled spleen protein samples from LDLR^−/−LMP7^−/− and LDLR^−/− mice at baseline and after feeding a high-fat Western-type diet (WD) over 6 weeks (n = 11 mice per group). LC indicates actin staining (for BMDM) or amidoblack staining (for spleen) as loading controls. (B) Representative Western blots of standard proteasome and immunoproteasome subunits of isolated 26 S proteasome derived from murine LDLR^−/−LMP7^−/− and LDLR^−/− BMDM (left panel) and spleen (right panel) lysates. (C) Chymotrypsin-, trypsin- and caspase-like proteasome activities (expressed as relative fluorescence units, RFU) of murine LDLR^−/−LMP7^−/− and LDLR^−/− BMDM; n = 4 experiments. Unpaired t-test or Mann-Whitney U test. Data in graphs are presented as mean ± SEM.