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. 2021 Nov 3;599(7886):657–661. doi: 10.1038/s41586-021-04062-5

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© The Author(s) 2021, corrected publication 2023

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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Extended Data Fig. 3 — a, Confocal microscopic images of tobacco leaf nuclei with expression of FRI–mScarlet I or FRL1–mScarlet I alone or co-expression of both. Data are representative of 5 independent experiments. b, c, Quantification of FRI–mScarlet I nuclear condensate area (b) and number per nucleus (c) in tobacco leaf with or without FRL1–mScarlet I co-expressed. An open circle indicates the median of the data and a vertical bar indicates the 95% confidence interval (CI) determined by bootstrapping. Numbers of nuclear condensates measured in (b) were 204 (FRI), 0 (FRL1) and 320 (FRI+FRL1). Numbers of nuclei analysed in (c) were 15 (FRI), 16 (FRL1) and 20 (FRI+FRL1). Comparison of mean by one-way ANOVA with adjustment (Sidak’s multiple comparisons test). ns, no significance. d, Confocal microscopic images of FRI–GFP nuclear condensates in the indicated mutants. e, f, Quantification of FRI–GFP nuclear condensate area (e) and number per nucleus (f) in the indicated genotypes. An open circle indicates the median of the data and a vertical bar indicates the 95% confidence interval (CI) determined by bootstrapping. Numbers of nuclear condensates measured in (e) were (from left to right) 391, 904, 159, 437, 324, 735, 354 and 448. Numbers of nuclei analysed in (f) were (from left to right) 181, 144, 96, 105, 132, 177, 142 and 126. At least 10 plants were analysed. Comparison of mean by one-way ANOVA with adjustment (Sidak’s multiple comparisons test). ns, no significance. g, Confocal microscopic images of tobacco leaf nuclei expressing FRI–mScarlet I and TAF15b–GFP. TAF15b–GFP was driven either by its endogenous protomer or overexpressed with CSV promoter. Data are representative of 3 independent experiments. h, Immunostaining images showing relative subnuclear localization of FRI–GFP (green) to U2B′′ (red) in root cells in NV and 2WV conditions. DNA was labelled with DAPI (blue). Non-tagged FRI was used a negative control. i, j, Quantification of the total number of U2B′′ condensates per nucleus (i) and those colocalized with FRI–GFP condensates (j) in NV and 2WV conditions. An open circle indicates the median of the data and a vertical bar indicates the 95% confidence interval (CI) determined by bootstrapping. n=166 nuclei (NV) and 130 nuclei (2WV). Comparison was via two-tailed t test with Welch’s correction. ns, no significance. In all the images, scale bars, 5 μm

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