# Fileset

[Supporting_information.pdf](https://mdr.nims.go.jp/filesets/c8026c7f-cf2c-4c16-93ed-e2ecb9fad4da/download)

## Creator

[Chiho Kataoka-Hamai](https://orcid.org/0000-0002-4068-0405), [Jingwen Song](https://orcid.org/0000-0003-1910-9287), [Kohsaku Kawakami](https://orcid.org/0000-0002-3466-9365)

## Rights

This document is the Accepted Manuscript version of a Published Work that appeared in final form in Langmuir, copyright © 2025 American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.langmuir.4c05063.[In Copyright](http://rightsstatements.org/vocab/InC/1.0/)

## Other metadata

[Incorporation of Triacylglycerol and Cholesteryl Ester Droplets in Phase-Separated Giant Unilamellar Vesicles](https://mdr.nims.go.jp/datasets/87e158fc-9cba-435f-a312-6a1b72e8b507)

## Fulltext

S1     Incorporation of Triacylglycerol and Cholesteryl Ester Droplets  in Phase-Separated Giant Unilamellar Vesicles  Chiho Kataoka-Hamai,*,1 Jingwen Song,1 and Kohsaku Kawakami1,2  1Research Center for Macromolecules and Biomaterials, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan 2Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan *E-mail: kataoka.chiho@nims.go.jp      SUPPORTING INFORMATION     Formation of Vesicles Consisting of a Single-Bilayer Region and a Double-Bilayer Region. We sometimes observed a vesicle consisting of a double-bilayer spherical segment and a single-bilayer spherical segment (Figure S1). We previously investigated such membrane structures obtained after the incorporation of triacylglycerol droplets in phosphatidylcholine vesicles in a single fluid phase labeled with Texas Red 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (TR-DHPE).1 In our previous study, the two membrane regions could be readily distinguished by their different TR-DHPE fluorescence intensities. However, in the present study, it was sometimes difficult to identify two-region vesicles. The vesicle bilayers consisted of liquid-ordered and liquid-disordered phases, which were labeled with different concentrations of TR-DHPE and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (ammonium salt) (NBD-DPPE) (Figure S1, magenta and green). Thus, it was not possible to determine the lamellarity based solely on the difference in the TR (or NBD) fluorescence intensities. Because of this difficulty, their membrane structures were not analyzed.S2          Reference (1) Kataoka-Hamai, C. Triacylglycerol-Droplet-Induced Bilayer Spontaneous Curvature in Giant Unilamellar Vesicles. Biophys. J. 2024, 123 (13), 1857–1868 FIGURE S1. (A) and (B) Vesicles consisting of double-bilayer and single-bilayer regions obtained after the incubation of DOPC/DPPC/cholesterol (3:3:2) GUVs labeled with 0.1% TR-DHPE (magenta) and 0.2% NBD-DPPE (green) with trieicosenoin droplets labeled with 0.005% TopFluor-TG (green). (i) Confocal fluorescence images and (ii) the corresponding membrane structures. Scale bars, 5 μm.