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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 5. — Vascular mural cell–specific deletion of SLIT3 reduces cardiac hypertrophy and dysfunction after transverse aortic constriction (TAC). A, Immunofluorescence staining of heart sections from Tbx18-CreERT2;Rosa26-Tdtomato mice using anti-SLIT3 antibody (green), anti-tdTomato antibody (red), and 4’,6-diamidino-2-phenylindole (DAPI; blue). White arrowheads denote the overlap of SLIT3 and tdTomato signals in the mural region of myocardial blood vessels in cross-section (left) and axially (right). Scale bar for (left), 10 µm. Scale bar for (right), 20 µm. B, Slit3 transcription determined by quantitative PCR (qPCR; normalized to Gapdh transcript levels) in Slit3^fl/fl or Tbx18-CreERT2;Slit3^fl/fl mice hearts after sham or TAC surgery. N=8 mice/group. C, Echocardiographic studies of Slit3^fl/fl and Tbx18-CreERT2;Slit3^fl/fl mice after sham or TAC surgery. N=8 mice/group. D, Appearance of hearts explanted from Slit3^fl/fl control and Tbx18-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 heart sections stained with wheat germ agglutinin (WGA; scale bar, 20 µm; bottom). E, Heart weight to body weight (HW/BW) ratio. N=8 mice/group. F, Quantification of cardiomyocyte cross-sectional area on WGA staining. N=600 cells from 6 mice in each group. G, qPCR analysis of hypertrophy-associated gene transcription (Nppa, Nppb, Myh7, and Acta1, normalized to Gapdh mRNA expression) in hearts from Slit3^fl/fl control or Tbx18-CreERT2, Slit3^fl/fl mice after sham or TAC surgery. N=8 mice/group. Two-way ANOVA with the Tukey multiple comparisons test was used in all analyses. EF indicates ejection fraction; LV, left ventricle; and LVPWd, end-diastolic left ventricle posterior wall thickness.