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. 2021 Oct 13;7(42):eabi6648. doi: 10.1126/sciadv.abi6648

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Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 5. — (A) Wheat germ agglutinin (WGA) immunofluorescence and trichrome staining of ECM deposition in heart sections from adult ctrl and knockout mice [ctrl, n = 2; dKO, n = 2; tKO (OR), n = 5; tKO (FR1), n = 5]. Scale bars, 20 μm. (B) Top 10 GO terms enriched in dKO compared to ctrl hearts. Terms associated with ECM are in red. (C) Microarray-based GSEA of adult ctrl and knockout hearts and heatmap of the 10 most up-regulated genes in dKO hearts derived from the respective gene set. Additional knockout of Osmr (tKO OR) or Fgfr1 (tKO FR1) or treatment with UO126 [ctrl: n = 4; dKO, dKO + UO, tKO (OR), tKO (FR1): n = 3] abolished enrichment in dKO hearts. (D and E) RT-qPCR analysis of different ECM genes in adult dKO (n = 4) and control cardiomyocytes (n = 5) and noncardiomyocytes (control, n = 4; dKO, n = 2 to 3) from 15-week-old mice, 3 weeks after tamoxifen administration. (F) Depiction of expression changes in neonatal and adult mice hearts of genes affecting ECM stiffness. (G and H) RT-qPCR analysis of agrin (F) (ctrl, n = 4; dKO, n = 3) and periostin (G) (ctrl, n = 4; dKO, n = 4) expression in isolated cardiomyocytes and noncardiomyocytes (n = 5) from adult dKO hearts compared to controls (two-tailed unpaired t tests). (I) Heatmap of genes related to soft ECM increased in miR-1/133a dKO versus wild-type (WT) hearts. Additional deletion of Osmr (tKO OR) or Fgfr1 (tKO FR1) or treatment with UO126 abolished increased expression. *P < 0.05 and ****P < 0.0001.