Figure 1.
Co-localized stains of serial sections of the aortic arch in Apo-E−/− mouse. Section thickness was set at 5 µm and original magnification at 100x.
A: Haematoxylin-Eosin stain for morphometry. B: Sirius red for collagen stain. C: Orcein for elastin stain. D: Immunohistochemistry, MMP-2 stain.
Figure 2.
Representative images from whole aorta sectioning and H+E staining in predefined levels (Descending Thoracic Aorta in four levels, Abdoninal Aorta above renal arteries in two levels and Abdominal Aorta below Renal Arteries in two levels.
Original magnification: 200x.
Figure 3.
Immunohistochemical staining with antibodies against MMP-8, MMP-9, TIMP-1, mac-3 and iNOS in optical microscopy sections of experimental groups CO, AT, EX and AT+EX.
Section thickness was set at 5 µm and original magnification at 100x.
Table 1.
Body-weight and plasma levels of glucose, total cholesterol, and triglycerides at the intervention start and the end of the study.
Table 2.
Atherosclerotic plaque area, lumen area, lumen stenosis (H&E staining), intra-plaque elastin and tunica media elastin content (orcein staining), intra-plaque collagen (sirius red staining) and total average intra-plaque MMP/TIMP positive stain (IHC).
Table 3.
Circulating concentrations of MMPs and TIMPs at the beginning and the end of the study (in ng/ml).
Table 4.
Densitometry measurements in zymography and reverse zymography gels.
Figure 4.
Verhoeff’s Van Gieson and Orcein stainings for the visualization of elastin content and stainings co-localization.
Lowercase letter image naming corresponds to a higher magnification of the defined section in the respectively named Uppercase named image. A–C: Verhoeff’s Van Gieson staining. D: Orcein staining. 1: Little or undetectable elastin. 2: Thick amorphous elastin content. 3: Formed elastin fibers. 4: Thin amorphous elastin.
Figure 5.
Morphometry and immunohistochemistry results for total plaque area in all examined aortic regions, collagen and elastin content, MMP, TIMP, mac-3, a-actin and iNOS intra-plaque localization and activity.
*: p value vs CO group <.05; ▴: ANOVA significance value <.05 and post hoc Tuckey HSD test p value vs AT group <.05; ▪: ANOVA significance value <.05 and post hoc Tuckey HSD test p value vs EX group <.05.
Figure 6.
Representative images from zymograms for the identification of metalloproteinase activity and reverse zymograms for the identification of TIMP activity.
A) Zymograhy, MMP-2, MMP-9 and proMMP-2 identification. B) Reverse zymography, TIMP-1 and TIMP-2 identification.