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. 2022 Mar 14;13:1310. doi: 10.1038/s41467-022-28833-4

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Cytoplasmic ROS (_cytROS) levels were measured using 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCF-DA) in apical areas of RHs in wild-type (Columbia Col-0), in the double mutant prx62-1 prx69-1 and in the 35S_proPRX62-tagRFP/Col-0 and 35S_proPRX69-tagRFP/Col-0 lines grown at 22 °C or 10 °C. Each point is the signal derived from a single RH tip. Fluorescence AU data are the mean ± SD (N = 18 root hairs), two-way ANOVA followed by a Tukey–Kramer test; (**) p < 0.01 (***) p < 0.001. Results are representative of two independent experiments. Asterisks indicate significant differences. NS = non-significant differences. Exact p-values are provided in the Source Data file. b Apoplastic ROS (_apoROS) levels were measured with Amplex™ UltraRed in apical areas of RHs in wild-type (Columbia Col-0), in the double mutant prx62-1 prx69-1 and in the 35S_proPRX62-tagRFP/Col-0 and 35S_proPRX69-tagRFP/Col-0 lines grown at 22 °C or 10 °C. Each point is the signal derived from a single RH tip. Fluorescence AU data are the mean ± SD (N = 11 root hairs), two-way ANOVA followed by a Tukey–Kramer test; (***) p < 0.001. Results are representative of two independent experiments. Asterisks indicate significant differences. Exact p-values are provided in the Source Data file. c Signal of SS-TOM and SS-EXT LONG-TOM in the apical zone of RHs grown at 22 °C or 10 °C with or without SHAM treatment in Col-0 and in the prx62-1 prx69-1 double mutant without SHAM. Cells were plasmolyzed with a mannitol 8% solution. In the images: (*) indicates cell surface including the plant cell walls, (**) indicates the retraction of the plasma membrane, (ap) apoplastic space delimitated between the plant cell wall and the retracted plasma membrane. Each point is the signal derived from a single RH tip. Fluorescence AU data are the mean ± SD SD (N = 11 root hairs), two-way ANOVA followed by a Tukey–Kramer test; (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Results are representative of two independent experiments. Asterisks on the graph indicate significant differences between each genotype 22 °C vs 10 °C. NS = non-significant differences. Exact p-values are provided in the Source Data file. Scale bars = 5 µm.