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. 2024 Feb 28;134(7):913–930. doi: 10.1161/CIRCRESAHA.122.321292

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© 2024 The Authors.

Circulation Research is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 3. — Fibroblast-specific deletion of SLIT3 reduces cardiac hypertrophy and dysfunction after transverse aortic constriction (TAC). A, Isolation of cardiac nonmyocyte (NCM) and cardiomyocyte (CM) cellular fractions from adult wild-type (WT) mice was verified by measuring Col1a1, Col3a1, Ccn2, Myh7, Myh6, and Acta1 mRNA levels (normalized to Gapdh transcript levels). Cells from N=4 WT mice and comparisons made with the Mann-Whitney U test. B, Slit3 transcript levels in isolated NCM and CM fractions from adult mouse hearts subjected to sham or TAC surgery. N=4 mice/group and comparisons were made with the Mann-Whitney U test. C, Immunofluorescence staining of a heart section from Tcf21-CreERT2;Rosa26-tdTomato mice using anti-SLIT3 antibody (green), anti-tdTomato (red), and 4’,6-diamidino-2-phenylindole (DAPI; blue). Scale bar, 50 µm. D, Slit3 transcript levels determined by quantitative PCR (qPCR) in Slit3^fl/fl and Tcf21-CreERT2;Slit3^fl/fl mice after 13 wk after sham or TAC surgery. N=8 mice/group. E, Echocardiography data from Slit3^fl/fl and Tcf21-CreERT2;Slit3^fl/fl mice after sham or TAC surgery with analysis of ejection fraction, fractional shortening, relative end-diastolic left ventricle posterior wall thickness (LVPWd), and relative left ventricle mass. N=8 mice/group. F, Explanted hearts from Slit3^fl/fl control or Tcf21-CreERT2;Slit3^fl/fl mice after sham or TAC surgery (top); representative images of heart sections stained with hematoxylin & eosin (H&E; scale bar, 500 µm; middle); and representative images of wheat germ agglutinin (WGA) stained heart sections (scale bar, 20 µm; bottom). G, Heart weight to body weight (HW/BW) ratio. N=8 mice/group. H, Quantification of CM cross-sectional area from WGA-stained sections. N=400 cells from 6 to 7 mice in each group. I, qPCR analysis hypertrophy-related genes (Nppa, Nppb, Myh7, and Acta1, normalized to Gapdh mRNA levels) in hearts from Slit3^fl/fl and Tcf21-CreERT2;Slit3^fl/fl mice after sham or TAC surgery. N=8 mice/group. Two-way ANOVA with the Tukey multiple comparisons test used in (D, E, and G–I).