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. 2024 Jul 16;15:5987. doi: 10.1038/s41467-024-50290-4

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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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Fig. 6 — a, b Co-IP assays indicate that ethylene impaired the interaction between OsETR2 (a) or OsERS2 (b) and OsCTR2, and weakened the interaction between MHZ3 and OsETR2 (a) or OsERS2 (b). Transgenic rice seedlings stably expressing OsETR2-Myc/WT and OsERS2-Myc/WT were grown in the dark for 2 days, followed by a 1-hour treatment with 10 μL/L ethylene. Total proteins were immunoprecipitated with anti-c-Myc affinity gel and immunoblotted with anti-c-Myc, anti-OsCTR2, anti-MHZ3, and anti-BiP antibodies. c Membrane association of OsCTR2 after 10 μL/L ethylene treatment in WT. Equal amounts of total protein (T), soluble protein (S), and microsomal membranes (M) were immunoblotted for OsCTR2, BiP (ER membrane marker), and UGPase (cytoplasm marker). d Luciferase complementation imaging assays indicate that ethylene attenuated the interaction between OsETR2 or OsERS2 and MHZ3. Equal amounts of the Agrobacteria harboring MHZ3-Nluc vector were co-infiltrated with Agrobacteria harboring either the Cluc-OsETR2 or Cluc-OsERS2 vectors into N. benthamiana leaves. After 48 hours at 22 °C, some plants underwent a 2-hour treatment with 10 μL/L ethylene, while others remained in ambient air. e The statistical analysis of the luminescence intensity in (d). The values represent the means ± SD, n = 7 biologically independent samples (**P < 0.01; two-tailed Student’s t-test; compared to the corresponding Air). f Immunoblot analysis for expression levels of Cluc-OsETR2, Cluc-OsERS2, and MHZ3-Nluc proteins in each experimental group. An unspecific band was utilized as an internal reference. g Ethylene (ET) promotes the interaction between OsEIN2 and MHZ3. Transgenic rice seedlings stably expressing OsEIN2-GFP were grown in the dark for 2 days, followed by a 1-hour treatment with 10 μL/L ethylene. Each experiment was repeated at least three times with similar results. Three independent experiments were repeated with similar results. Uncropped blots and source data are in the Source Data file.