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. 2022 Jul 26;23(8):1169–1182. doi: 10.1038/s41590-022-01272-5

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© The Author(s) 2022

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. 3 — a,b, Actomyosin structure in FRCs day 3 (a) and day 5 (b) after IFA/OVA immunization for perlecan (matrix, magenta), PDPN (FRC, yellow), phalloidin (F-actin, cyan), and pMLC (red). Asterisks and arrowheads indicate F-actin cables with or without perlecan alignment. Orthogonal views (yellow dotted line, yz axis, 10 μm depth). Representative image of three independent experiments. Scale bars, 10 µm. c, Percentage of Fibers. Stacked bar plots with mean ± standard deviation (s.d.) error bars. d, Confetti-labeled FRCs (yellow) spanning space without (white asterisk) ECM (magenta). Scale bar, 5 µm. e, Percentage of Fibers. Box plot indicates median, interquartile range, and minimum/maximum. One-way ANOVA with Tukey’s multiple comparisons, **P < 0.01. c,e, n indicates image ROI on day 0 (n = 14), day 3 (n = 13), and day 5 (n = 12) from three mice. f, Initial recoil velocity (µm s⁻¹) after IFA/OVA immunization, ± ROCK inhibition (Y27632). Resistive myosin forces (blue text indicates the ratio between PBS/Y27). Box plot indicates median, interquartile range, and minimum/maximum. Two-way ANOVA with Tukey’s multiple comparisons, ****P < 0.0001, **P = 0.001973. g, Recoil curves of network displacement (µm) (mean ± s.e.m.). f,g, n indicates an ablation on day 0 (n = 54), day 0 + Y27 (n = 43), day 3 (n = 18), day 3 + Y27 (n = 20), day 5 (n = 19), and day 5 + Y27 (n = 15) over three independent experiments. h, Actomyosin structure in FRCs day 5 after IFA/OVA immunization with or without ROCK inhibition (Y27632). Perlecan (matrix, magenta), PDPN (FRC, yellow), phalloidin (F-actin, cyan), and pMLC (red). Asterisks and arrowheads indicate F-actin cables with or without perlecan alignment. Representative image of three independent experiments. Scale bars, 10 µm. i, Percentage of Fibers. Stacked bar plots with mean ± s.d. error bars. Two-way ANOVA with Tukey’s multiple comparisons, ****P = 0.000577, *P = 0.03824. j, Percentage of Fibers. Box plot indicates median, interquartile range, and minimum/maximum. One-way ANOVA with Tukey’s multiple comparisons, ****P < 0.0001. i,j, n indicates image ROI on day 0 (n = 12), day 0 + Y27 (n = 12), day 5 (n = 18), and day 5 + Y27 (n = 18) from three mice. NS, not significant.

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