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. 2023 Oct 10;12(1):2268261. doi: 10.1080/21623945.2023.2268261

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© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 2. — Hydrogel density regulates lipid droplet (LD) size independent of cell size or LD number.

a. Cell area (μm2) of VAT or SAT mature adipocytes differentiated for 14 d in 3%, 5%, or 10% PEG-VS gels determined by Phalloidin staining and measured using FIJI. Each data-point represents the mean cell area (μm2). No statistically significant differences were observed between experimental arms. b. Number of lipid droplets per mature adipocyte in 3%, 5%, or 10% PEG-VS gels determined by LipidTox staining and measured using FIJI. Each data-point represents the mean number of LD per cell. No statistically significant differences were observed between experimental arms. c. Mean lipid droplet area in mature adipocyte pooled from 3%, 5%, and 10% PEG-VS gels, separated by VAT and SAT. Each data-point represents the mean LD area per cell. Statistically significant differences between experimental arms denoted by p-values over indicated bars (ANOVA). d. Mean lipid droplet area in mature adipocyte in 3%, 5%, or 10% PEG-VS gels and separated by VAT and SAT. Each data-point represents the mean LD area per cell. Statistically significant differences between experimental arms denoted by p-values over indicated bars (ANOVA, Tukey’s multiple comparisons test). e. Total lipid droplet area/cell area as a measure of lipid density in adipocytes, in 3%, 5%, or 10% PEG-VS gels, separated by VAT and SAT. Statistically significant differences between experimental arms denoted by p-values over indicated bars (ANOVA, Tukey’s multiple comparison test). f. Representative confocal microscopy images of mature adipocytes in PEG-VS gels. g, h. Size frequency distribution of lipid droplet area in adipocytes differentiated for 14 d in 3%, 5%, or 10% PEG-VS gels quantified using confocal microscopy and FIJI. Small, Medium, Large, and X-Large droplets defined as <9.2 mM2, 9.2-14.5 mM2, 14.6-18.6 mM2, >18.6 mM2, respectively, for VAT, and <9.8 mM2, 9.8-14.7 mM2, 14.8-22.6 mM2, >22.6 mM2, respectively, for SAT. Each data-point represents the relative frequency of that size LD per cell. Statistically significant differences between experimental arms denoted by p-values over indicated bars (ANOVA, Tukey’s multiple comparison test). Data from 7 to 8 cells per patient from four patients, 2 M, 2F.