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. 2024 Mar 29;15:2766. doi: 10.1038/s41467-024-46774-y

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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. 2 — a Representative 3D reconstructions of hydrogels containing various GH content (0, 1, 3 wt%), revealing bicontinuous structures in 1 and 3 wt% hydrogels (gelatin-rich (GR, green) and gelatin-poor (GP, gray) domains). Scale bar = 50 μm. b Fraction of total volume occupied by GR (left panel) and GP (right panel) domains within hydrogels. n = 9 (0%), 8 (1%), or 7 (3%) regions across 3 hydrogels per condition. Left panel: 0% vs. 1%, 1% vs. 3% ****p ≤ 0.0001. Right panel: 0% vs. 1%, 0% vs. 3% ****p ≤ 0.0001; 1% vs. 3% *p = 0.0353; one-way ANOVA with Tukey post hoc. c Quantification of connectivity (defined by percent of total volume occupied by largest individual domain) of GR (top) and GP (bottom) domains. n = 9 (0%), 8 (1%), or 7 (3%) regions across 3 hydrogels per condition. Upper panel: n.s. indicates no statistical significance. Lower panel: 0% vs. 1%, 0% vs. 3% ****p ≤ 0.0001; one-way ANOVA with Tukey post hoc. d Quantification of interfacial surface area normalized to sample volume across hydrogels containing various GH content (0, 1, 3 wt%). n = 9 (0, 3%) or 8 (1%) regions across 3 hydrogels per condition. 0 % vs. 1%, 0% vs. 3%, 1% vs. 3% ****p ≤ 0.0001; one-way ANOVA with Tukey post hoc. e Representative elastic modulus maps determined with AFM-nanoindentation (empty nodes denote removed outliers). f Average elastic moduli (top) and coefficient of variation (CV, bottom) across AFM profiles. n = 9 (0%), 7 (1%), 8 (3%) grids across 3-4 hydrogels per condition. Upper panel: 0% vs. 1% **p = 0.0041; 0% vs. 3% **p = 0.0014. Lower panel: 0% vs. 1% *p = 0.0354; 0% vs. 3% *p = 0.0228; one-way ANOVA with Tukey post hoc. g Quantification of averaged elastic moduli of GR and GP domains across probed grids for 1 wt% GH hydrogel using AFM based on differential fluorescent labeling (individual data points in Supplementary Fig. 7d). n = 8 (GP) or 11 (GR) grids across 5 hydrogels. *p = 0.0333; Two-tailed unpaired students t-test. Data are mean ± s.d. Source data for (b–g) provided as a source data file.