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. 2023 Jan 24;14:379. doi: 10.1038/s41467-023-36046-6

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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. 1 — a Schematic illustration of the optoDroplet assay in HEK293 cells. Candidate genes were fused with mCherry and Cry2. The RNA-binding protein FUS was chosen as a positive control. Unfused mCherry-Cry2 was used as a negative control. Upon blue light exposure, proteins with phase-separation capacity formed condensates in cells. b OptoDroplet assay of Rph3A performed in HEK293 cells. The number of puncta per cell was counted and plotted. Data are displayed as the mean ± SEM (n = 6 cells per group). c Representative images and quantification of fluorescence recovery from the FRAP analysis of Rph3A droplets. The time point of bleaching was 0 s. The data are displayed as the mean ± SEM (n = 6 droplets). d Fusion events of droplets composed of Rph3A-mCherry-Cry2 in HEK293 cells captured by time-series imaging. e Rph3A-mCherry-Cry2 droplets condensed with Rph3A-EGFP after blue light stimulation. FUS and EGFP were substituted for Rph3A as negative controls. f Magnified images, in which the lines indicate the fluorescence intensity profiles of puncta in the white squares in e. Scale bar, 1 μm. g Representative images and quantification of fluorescence recovery from the FRAP analysis of Rph3A-EGFP droplets. The data are displayed as the mean ± SEM (n = 5 droplets). h Fusion events of Rph3A-EGFP and Rph3A-mCherry-Cry2 condensates in HEK293 cells stimulated with blue light. The image of cell at time point 0 s (left) and the magnified time-lapse images of the white squares in left (right) are showed. Source data and of b, c and g are provided in the Source Data file.