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. 2021 Nov 29;36(4):994–1005. doi: 10.1038/s41375-021-01432-w

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© The Author(s) 2021

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. 2 — A MIP vector and MIP-OVA retrovirus constructs used in generation of C1498-MIP and C1498-OVA cell lines. The murine Cadm1 signal peptide (SP) and transmembrane domain (TMD) were cloned 5’ to full-length chicken ovalbumin into the MIP vector. B Representative flow cytometry plots and histogram of C1498-MIP and C1498-OVA cell lines stained with antibodies against MHC class I-presented OVA peptide 257–264 (H-2K^b:SIINFEKL), which demonstrate OVA antigen presentation in C1498-OVA cells compared to C1498-MIP or fluorescence minus one (FMO) negative control staining. C OVA-specific CD8⁺ T cell (B3Z) lacZ activation assay. C1498-MIP or C1498-OVA cells were incubated with B3Z CD8⁺ T cell hybridoma reporter cells, in which OVA-specific T cell receptor activation drives lacZ expression. Representative image demonstrating OVA-specific T cell activation (red color) in B3Z lysates, as assayed with the β-gal substrate chlorophenol red-β-galactoside (CPRG).