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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 1. — Inducible global-SLIT3 deficiency alleviates the development of cardiac hypertrophy and dysfunction after transverse aortic constriction (TAC). A, SLIT3 transcription as measured by RNA sequencing and displayed as reads per kilobase of transcript per million mapped reads (RPKM) in ventricular tissue samples from normal controls and patients with pressure overload induced by congenital heart disease. Each data point represents a unique subject. Comparison performed with Mann-Whitney U test. B, Immunofluorescence staining of myocardial sections Slit3^fl/fl and Rosa26-CreERT2;Slit3^fl/fl mice 4 wk after tamoxifen injection using anti-SLIT3 antibody (green) and DAPI (blue) demonstrated loss of SLIT3 in the Rosa26-CreERT2;Slit3^fl/fl mice. Scale bar, 500 µm. C, TAC experimental design and timeline. D, Slit3 transcript levels in hearts from Slit3^fl/fl and Rosa26-CreERT2;Slit3^fl/fl mice after sham or TAC surgery. E, B-mode and M-mode echocardiography in Slit3^fl/fl and Rosa26-CreERT2; Slit3^fl/fl mice after sham or TAC surgery with analysis of ejection fraction (EF), shortening fraction (FS), relative left ventricle posterior wall thickness at diastole normalized to mice body weight (end-diastolic left ventricle posterior wall thickness [LVPWd]), relative left ventricle mass, and normalized to body weight. N=8 mice/group. F, Explanted hearts from Slit3^fl/fl and Rosa26-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) staining of myocardial sections (scale bar, 20 µm, bottom). G, Heart weight to body weight (HW/BW) ratio in Slit3^fl/fl or Rosa26-CreERT2;Slit3^fl/fl mice after sham or TAC surgery. N=8 mice in each group. H, Quantification of myocyte cross-sectional area from WGA staining. N=400 cells from 6 to 8 mice in each group. I, Quantitative PCR (qPCR) analysis of transcript levels of hypertrophy-associated genes (Nppa, Nppb, Myh7, and Acta1, normalized to Gapdh transcript levels) in hearts from Slit3^fl/fl and Rosa26-CreERT2;Slit3^fl/fl mice after sham or TAC surgery. N=8 mice in each group. Two-way ANOVA with the Tukey multiple comparisons test used in (D through I). DAPI indicates 4’,6-diamidino-2-phenylindole.