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. 2024 Apr 24;21(9):1702–1707. doi: 10.1038/s41592-024-02242-5

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© The Author(s) 2024

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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Extended Data Fig. 5 — 2-plex Exchange-PAINT image of transiently transfected CHO cells expressing CD86-ALFA-mEGFP on the cell surface. Respective anti-CD86 antibody was site-specifically modified via either transglutaminase a, or via GlyCLICK b, DNA-strand conjugation. Schematic sketch of labeling efficiency determination for target anti-CD86 antibody (cyan) specifically binding the membrane receptor CD86. For antibody characterization ALFA-tag was inserted at the CD86 N-terminus on the intracellular side of the cell surface and the fluorescent protein mEGFP served as an indicator for surface expression. Colocalization of target antibody and ALFA-tag nanobody reference signal (magenta) revealed underlying labeling efficiency. Zoom-ins depict super-resolution representation of target anti-CD86 antibody colocalizing with respective ALFA-tag nanobody.