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. 2021 Jun 24;12:3928. doi: 10.1038/s41467-021-24215-4

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© The Author(s) 2021

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. 1 — a Western blotting for Thbs1 in heart tissue of mice (8–10 weeks of age) subjected to 2 weeks of TAC, that contained the activated calcineurin A transgene (ΔCnA), or are Csrp3^−/−, compared to sham-operated or wild-type (WT) control. Gapdh is shown as a processing and loading control. b Representative immunohistochemistry for endogenous Thbs1 (red) and cell outlines with wheat germ agglutinin (WGA)-FITC (green) with DAPI-stained nuclei (blue) from hypertrophic ΔCnA transgenic and WT control hearts at 8 weeks of age. Scale bars are 10 μm. c Immunohistochemistry for Thbs1 protein (green), vimentin (red) from sham or TAC-operated hearts, 2 weeks later. Scale bars are 50 μm. Nuclei are shown in blue with DAPI. d Schematic diagram depicting the inducible double transgenic (DTG) tetracycline-repressor system for inducible overexpression of Thbs1 in the heart. e Representative Western blots for Thbs1, Thbs3, Armet, BiP, calreticulin (calret.), and Gapdh as a loading control from hearts of tTA cont., Thbs1 DTG, and Thbs3 DTG mice at 6 weeks of age. f Representative immunohistochemistry broken into 2 channels each for overexpressed Thbs1 (green) with WGA-labeled membranes (purple), DAPI for nuclei (blue) and BiP (red) to show ER and the vesicular compartment in Thbs1 DTG hearts at 8 weeks of age. Scale bars are 50 μm. g Representative images of transmission electron microscopy of heart sections from tTA cont. and Thbs1 DTG mice at 6 weeks of age. Upper panels: arrowheads indicate ER in tTA cont., white arrows show expanded ER and vesicles only in Thbs1 DTG hearts. Lower panels: enlargement of white dotted boxed area from upper panels. Scale bars are 1 μm. Source data are provided as a Source Data File.