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. 2024 Dec 30;15:10910. doi: 10.1038/s41467-024-55292-w

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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. 6 — a The 5-day-old EYFP-ATG8f/WT and EYFP-ATG8f/ubp1abc plants were subjected to HS treatment at 38 °C for 1 h, and then recovered at 22 °C for 6 h, 9 h, and 12 h. The maturation zone of indicated samples was observed using a confocal microscope. Scale bar = 10 μm. The magenta triangle represents circular structures. b Statistical analysis of the number of punctate signals in (a). Data represent mean ± SD; n = 5. Statistical analysis was performed using a two-tailed unpaired Student’s t test. c The 5-day-old EYFP-ATG8f/WT and EYFP-ATG8f/ubp1abc plants were subjected to HS treatment at 38 °C HS for 1 h, and then recovered in 1/2 MS liquid medium supplemented with 1 μM Conc A for 6 h, 9 h, and 12 h at 22 °C. The maturation zone of the roots from the indicated samples was observed using a confocal microscope. Scale bar = 10 μm. d Statistical analysis of the number of autophagosomes in (c). Data represent mean ± SD; n = 15. Statistical analysis was performed using a two-tailed unpaired Student’s t test. e The 5-day-old WT and ubp1abc plants were subjected to treatment at 22 °C (control) or 38 °C (HS) for 1 h and then heat-stressed plants recovered at 22 °C for 6 h, 9 h, and 12 h. Total proteins were extracted to detect using Anti-NBR1 (Agrisera; Cat# AS14 2805 A; 1:3000) and Anti-ATG8 (Agrisera; Cat# AS14 2769; 1:3000), with Anti-actin (Sangon Biotech; Cat# D110007; 1:5000) as the internal control. The value represents the relative protein content, defining the control group as 1. Data represent mean ± SEM; n = 3. f The 5-day-old WT and ubp1abc plants were subjected to treatment at 22 °C (control) or 38 °C (HS) for 1 h and then heat-stressed plants recovered at 22 °C for 6 h and 12 h. The same amount of insoluble protein was separated by SDS-PAGE and then stained with silver stain, which was used as a loading control. g Statistical analysis of the insoluble protein content in (f). Data represent the mean ± SD; n = 3. Statistical analyses were performed using a two-tailed paired Student’s t test. Source data are provided as a Source Data file.