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. 2021 Sep 6;12:5270. doi: 10.1038/s41467-021-25653-w

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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. 3 — a Western blot analysis showing Nrf1 expression in hearts from mice at 4-week after AAV9-TdTomato and AAV9-Nrf1 injection at P4; experiments were repeated independently twice with similar results. b Illustration of a timeline showing the experimental design for AAV9 injection and time points of sample collections. c TTC staining showing infarct size on transverse heart sections collected at the end of reperfusion; Scale bar, 500 μm. d Quantification of infarct area in sections from (c); AAV9-TdTomato, n = 6; AAV9-Nrf1, n = 5; *p = 0.0183, by Student’s t-test two-tailed. e–g Fractional shortening (e), ejection fraction (f), and systolic volume (g) of left ventricles from AAV9-TdTomato and AAV9-Nrf1 hearts at baseline (before I/R), and 1–3 weeks (wk) after I/R; n = 5 animals for each group. e 2wk, *p = 0.0388; 3wk, *p = 0.034. f 2wk, *p = 0.0422; 3wk, *p = 0.0336. g 1wk, *p = 0.0207, 2wk, *p = 0.0188; 3wk, *p = 0.0155. h Masson’s trichrome staining showing fibrotic scarring on transverse sections of hearts collected at 600 μm below the ligation suture; Scale bar, 500 μm. i Quantification of fibrotic regions of heart sections collected at multiple planes below the ligation suture from AAV-TdTomato and AAV-Nrf1 mice; AAV9-TdTomato, n = 5; AAV9-Nrf1, n = 4; 200 μm, *p = 0.012, 400 μm, *p = 0.0455, 600 μm, *p = 0.0007, 800 μm, *p = 0.0012. j Viable NRVMs with adenoviral (Ad) overexpression of LacZ or Nrf1 following H₂O₂, peroxynitrite, doxorubicin, and erastin treatments, indicated by Calcein AM (green); Scale bar, 100 μm. k Cell death measured by lactate dehydrogenase (LDH) release in NRVMs-overexpressing LacZ, Nrf1, or Nrf2 following toxin treatments; n = 4 biologically independent experiments for each group. H₂O₂, ***p < 0.000001, **p = 0.0043. Peroxynitrite, ***p < 0.000001, **p = 0.0022. Doxorubicin, ***p = 0.000006, **p = 0.0003. Erastin, **p = 0.0039, *p = 0.0215. d–g, i, k Results are shown as mean ± s.e.m.; Student’s t-test two-tailed; n.s. not significant.