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. 2023 Jan 4;55(1):108–119. doi: 10.1038/s12276-022-00918-6

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© The Author(s) 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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Fig. 4 — a Representative fluorescence images of COS7 cells transfected with EGFP-nArgBP2_959-1196 and SBFP2 empty vector or CaMKIIα-SBFP2. EGFP-nArgBP2_959-1196 droplets coexpressed with CaMKIIα-SBFP2 dispersed upon ionomycin treatment in 4 mM Ca²⁺ Tyrode (middle), while no effect on the droplets was observed without CaMKIIα coexpression (top) or in the absence of extracellular Ca²⁺ (bottom). Scale bar: 20 μm. b Enlarged time-lapse images of EGFP-nArgBP2_959-1196 droplets coexpressed with CaMKIIα-SBFP2 in the middle panel of Fig. 4a before and after ionomycin treatment. Scale bar: 1 μm. c Plot of the normalized average fluorescence intensity profiles of EGFP-nArgBP2_959-1196 droplets (filled circle) and the surrounding cytosol (open circle) over time. n = 54 for both droplets and surroundings. Values are means ± s.d. d Schematic diagram of nArgBP2_959-1196-3S3D (phospho-mimetic) and nArgBP2_959-1196-3S3A (phospho-deficient) mutants. e Representative fluorescence images of COS7 cells transfected with EGFP-nArgBP2_959-1196-3S3D and EGFP-nArgBP2_959-1196-3S3A. Scale bar: 20 μm. f Representative fluorescence images of COS7 cells cotransfected with EGFP-nArgBP2_959-1196-3S3A and CaMKIIα-SBFP2 and treated with ionomycin or KN93 and ionomycin in the presence of 4 mM Ca²⁺. Scale bar: 20 μm. g Fluorescence images of purified EGFP-nArgBP2_959-1196 before treatment with CaMKIIα and ATP. h Enlarged time-lapse images of purified EGFP-nArgBP2_959-1196 (yellow squares in g) before and after treatment with CaMKIIα and ATP. Scale bar: 2 μm. i Line scan plots of EGFP-nArgBP2_959-1196 (green dashed line in h) fluorescence intensity before and after CaMKIIα and ATP treatment.