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. 2024 Nov 1;15:9448. doi: 10.1038/s41467-024-53894-y

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 7 — a, b Relative mRNA level of PR1a (a) and PR2 (b) in wild type and CMV 2b-transgenic N. benthamiana plants. c, d Relative mRNA level of PR1a (c) and PR2 (d) in wild type and TuMV HC-Pro-transgenic N. benthamiana plants. e, f Interactions between CMV 2b (e) or TuMV HC-Pro (f) with NbHSP90s in a yeast two-hybrid assay. Yeast strain AH109 transformed with the indicated plasmid combinations was spotted with 10-fold serial dilutions on synthetic dextrose media SD/-Leu-Trp and SD/-Ade-His-Leu-Trp. g, h Interaction between CMV 2b and NbHSP90-2 (g) or NbHSP90-10 (h) in the co-IP assay. CMV 2b-Flag and NbHSP90-2-HA were co-expressed and CMV 2b-Flag+GFP-HA served as a negative control. i, j Interaction between TuMV HC-Pro and NbHSP90-2 (i) or or NbHSP90-10 (j) in the co-IP assay. k, l Effect of CMV 2b (k) and TuMV HC-Pro (l) on the SA-induced degradation of NbNPR3-Flag in the semi-in vivo assays. NbNPR3-Flag was co-expressed with CMV 2b-Flag (or TuMV HC-Pro) or GUS-Flag (control). Protein extracts were mixed with CHX, ATP and SA and then subjected to semi-in vivo protein degradation. NbNPR3-Flag protein was analyzed with anti-Flag antibodies at different time points. Ponceau S staining was performed to determine the amount of protein load. N = 6–10 samples (2–3 plants per sample) for (a, b), 5–7 samples (2–3 plants per sample) for (c, d). Data are mean ± SEM. n. s. stands for no significant difference, *P < 0.05, and **P < 0.01 (one-way ANOVA for a–d). Source data and the exact P values are provided in Source Data file.