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. 2024 May 15;629(8014):1126–1132. doi: 10.1038/s41586-024-07424-x

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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. 6 — a, Fluorescence of mCherry-tagged JAM2 and JAM3 in yeast nuclei at 20 °C (Top) and 30 °C (Bottom). Scale bar 5 µm. b, FRET-FLIM analysis of GFP-TWA1 with mCherry-tagged acceptor proteins JAM2, JAM3, or no acceptor (−) at 30 °C in the yeast nucleus. Reduction of TWA1 fluorescence lifetime by JAM2 expression indicates proximity of the proteins which occurred within nuclear subdomains (see a). Asterisks indicate a significant difference (JAM2, nucleus, P = 0.009; JAM2, inside subdomain, P = 0.008; two-sided Mann-Whitney U test, 20-30 cells). c, Protein interaction analysis of mCherry-tagged JAM2 with GFP and GFP-twa1 product (twa1). The analysis with non-fused mCherry protein is shown as a control (−). Two-sided Mann-Whitney U-Test, P = 0.82 (JAM2, twa1); P = 0.68 (mCherry, GFP); n = 20−30. d, No detectable interaction of TWA1 with JAM2 at 20 °C in yeast nucleus. FRET-FLIM analysis with GFP-TWA1 (−) and GFP-TWA1 with mCherry-tagged acceptor proteins JAM2 and JAM3 (two-sided Mann-Whitney U-Test, P = 0.24 (JAM2, TWA1); P = 0.07 (JAM3, TWA1); n = 20−30. Box plots as in SI Extended Data Fig. 5. Statistical significance in SI_Extended Fig. 6.

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