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. 2022 Jul 19;9:770065. doi: 10.3389/fcvm.2022.770065

FIGURE 3.

FIGURE 3

Effect of Tgfb1 and Smad3 heterozygous deletion on collagen and elastin fibers in the aortic valve of 10–12-week-old Kl–/– mice. (A–H) Verhoeff-Van Gieson (VVG) staining of tissue sections through the aortic valve and annulus of wild-type (A,E), Kl–/– (B,F), Kl–/–;Tgfb1± (C,G), and Kl–/–;Smad3± (D,H) mice. Magnified images of the boxed area (A–D) are given in (E–H). Arrows indicate the presence of abnormal elastin fibers in Kl–/– (F) and Kl–/–;Tgfb1± (G) mice. VVG stains elastin in black and collagen fibers in red color. (I) Histogram showing total collagen content in the aortic valve, including leaflets, annulus, and AV hinge. Quantification of collagen fibers content was done by measuring the intensity of collagen staining on multiple serial sections from each animal per group by using NIH-Image J software. Values indicate mean ± SD (n = 5). (J) Histogram showing the average area containing the elastin fibers in the AV hinge. Quantification of elastin fibers content was done by measuring the average area in the AV hinge with elastin staining on multiple serial sections from each animal per group by using NIH-Image J software. Values indicate mean ± SD (n = 5), and significant “p-values” were given on the top of histograms. Scale bars: (A–H) 200 μm.