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. 2023 Feb 23;72(7):2113–2125. doi: 10.1007/s00262-023-03409-3

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© The Author(s) 2023

Open AccessThis 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. 2 — Linker sequences module antigen presentation. a Schematic representation of the constructs used. Gray: signal peptide (SP); orange: FLAG tag; blue: neoantigens; green: SIINFEKL peptide; and, yellow: mouse CD44 transmembrane domain. b Graphical scheme of the experimental procedure used for linker screening. c Western blot analysis of the expression of recombinant proteins untreated transfected B16-F10 cells. Asterisk indicates expected molecular weight. d Western blot analysis of the expression of recombinant proteins in IFN-g stimulated transfected B16-F10 cells. Asterisk indicates expected molecular weight. e Western blot analysis of the expression of recombinant proteins in MG132-treated transfected B16-F10 cells. Asterisk indicates expected molecular weight. f Western blot analysis of the expression of recombinant proteins in IFN-g stimulated and MG132-treated transfected B16-F10 cells. Asterisk indicates expected molecular weight. g Representative flow cytometry panels for the detection of MHC-I/SIINFEKL on the surface of transfected B16-F10 cells. Frequency of positive cells for the linker under examination is represented on each panel. Gray: mock; green: linker under examination; purple: SIINFEKL positive control. h Levels of MHC-I/SIINFEKL on the surface of B16-F10 transfected cells expressed as geometric mean plus SD of three independent experiments. i Frequency of MHC-I/SIINFEKL positive cells. Mean plus SD of three independent experiments is shown. Data were analyzed using Dunn’s test, ^**P < 0.01, ^*P < 0.1