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. 2024 Aug 27;15:7397. doi: 10.1038/s41467-024-51299-5

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© The Author(s) 2024

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

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Fig. 5 — a A reaction landscape of the division of ternary-mixed DNA droplets. The division pathway indicates the order of droplet division. b Formation of ternary-mixed DNA droplet (C·A·B-droplet) containing three types of Y-shaped DNA nanostructures and two types of DNA linkers. c Schematic of pathway-controlled division of C·A·B-droplet. The linker-cleavage reaction rates decide the order of droplet division, thereby changing the pathway of droplet division. d Time-lapse images of C·A·B-droplet division in Pathway 1 before and after adding T^†_ACi, iT^†_ABi, R^†_ABi, and RNase H. The detail of multistep division process is shown in Supplementary Movie 6. R^†_ABi (i = 1, 2): miR-6875-5p and miR-4634. Scale bars: 20 μm. e Time-lapse images of C·A·B-droplet division in Pathway 2 before and after adding T^†_ABi, iT^†_ACi, R^†_ACi, and RNase H. The detail of multistep division process is shown in Supplementary Movie 7. R^†_ACi (i = 1,2): miR-1246 and miR-1307-3p. Scale bars: 20 μm. f, g Time courses of division ratio r_{div_B} (blue) and r_{div_C} (red) during C·A·B-droplet division in Pathway 1 (f) and Pathway 2 (g), respectively. Data are presented as the mean ± standard error (SE) of three field of view of microscopy observation.