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. 2022 Jul 21;18(10):1104–1114. doi: 10.1038/s41589-022-01062-y

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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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Extended Data Fig. 7 — (a) Model of NPM1 with the identified phosphosites highlighted in orange. Protein domains: OD- oligomerization domain, ABP- acidic basic patch and NBD- nucleic acid binding domain. (b) Scatter plot of calculated partition coefficient (K) from SiR-SNAP channel (y-axis) and size of individual (segmented-) nucleoli (x-axis) from at least three independent trials per construct. (c) Scatter plot of calculated partition coefficient from SiR-SNAP channel (K_SiR-SNAP, y-axis) and mean intensity of SiR-SNAP in nucleoli (proxy for the expression level of SNAP-tagged protein, x-axis) for individual (segmented-) nucleoli from at least three independent trials per construct. (d) Box plot showing the distribution of K_SiR-SNAP (partition coefficient calculated from SiR-SNAP channels for NPM1 wildtype (wt) and phosphomutants. The box plots display the median and IQR, with the upper whiskers extending to the largest value ≤1.5 × IQR from 75th percentile and the lower whiskers extending to smallest values ≤1.5 × IQR from 25th percentile. (e) Scatter plot of calculated partition coefficient from GFP channel (K_GFP, y-axis) and mean intensity of GFP in nucleoli (proxy for the expression level of GFP-tagged wt-NPM1, x-axis) for individual (segmented-) nucleoli from at least three independent trials per construct. (f) Box plot showing the distribution of K_GFP (partition coefficient calculated from GFP channels for NPM1 wildtype (wt) expressed as marker of nucleoli in all experiments. The box plots display the median and IQR, with the upper whiskers extending to the largest value ≤1.5 × IQR from 75th percentile and the lower whiskers extending to smallest values ≤1.5 × IQR from 25th percentile. (g) Barplot representing the relative protein abundance (in log₂ scale, y-axis) of heterologously expressed SNAP-tagged phosphomutants (x-axis) of NPM1 compared to the wild type (wt) from three biological replicates.

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