Ldlr-KO(2)

Fig.1 The body weight curve of Ldlr-KO mice and WT mice under CD conditions (n=4-10).

Abbr. WT, wild type; HO, homozygous; CD, chow diet .

Fig 2. Blood lipid profile of Ldlr-KO mice and WT mice under CD conditions (n=6-10, Mean±SEM, Unpaired t-test, *P < 0.05, **P< 0.01，***P < 0.001).

Abbr. WT, wild type; HO, homozygous; CD, chow diet.

Fig 3. Oil Red O staining of aortic valve in male Ldlr-KO mice and WT mice under CD conditions. 

The results indicated that no lipid accumulation was observed in the aortic valves of male WT mice at 7 months of age, while all male Ldlr-KO mice (2/2) exhibited lipid accumulation in the aortic valves at 3 months of age, with the severity of this phenotype progressively worsening with age. (n=2).

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 4. Oil Red O staining of aortic valve in female Ldlr-KO mice and WT mice under CD conditions. 

The results indicated that no lipid accumulation was observed in the aortic valves of female WT mice at 7 months of age, while all female Ldlr-KO mice (2/2) exhibited lipid accumulation in the aortic valves at 3 months of age , with the severity of this phenotype progressively worsening with age. (n=2).

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 5. Oil Red O staining of aorta in male Ldlr-KO mice and WT mice under CD conditions (n=2). 

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 6. Oil Red O staining of aorta in female Ldlr-KO mice and WT mice under CD conditions (n=2). 

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 7. Masson staining of aortic valve in male Ldlr-KO mice and WT mice under CD conditions. 

Muscle fibers appear red, while collagen fibers are stained blue. Ldlr-KO mice exhibited increased collagen content within plaques, suggesting enhanced plaque stability (n=2). 

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 8. Masson staining of aortic valve in female Ldlr-KO mice and WT mice under CD conditions. 

Muscle fibers appear red, while collagen fibers are stained blue. Ldlr-KO mice exhibited increased collagen content within plaques, suggesting enhanced plaque stability (n=2). 

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 9. Representative pictures of aortic valve. 

In male WT mice, no significant plaque formation, necrosis, or inflammatory cell infiltration was observed. However, in male Ldlr-KO mice, slight proliferation of connective tissue were seen in the luminal surface (black arrow), along with small amount of infiltration of foam cells (blue arrows). A small amount of necrotic tissue was noted (red arrows). Myocardial cells displayed loose and irregular arrangement. The black box indicates the location of the magnified view.

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 10. Representative pictures of aortic valve. 

In female WT mice, no significant infiltration of foam cells were observed. However, in female Ldlr-KO mice, a small amount of infiltration of foam cells were seen in vascular intima (blue arrows). Myocardial cells displayed loose and irregular arrangement. The black box indicates the location of the magnified view.

Abbr. WT, wild type; HO, homozygous; CD, chow diet; M: month.

Fig 11. The body weight curve of Ldlr-KO mice and WT mice under HFD conditions. (n=4-10). 

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 12. Blood lipid profile of Ldlr-KO mice and WT mice under HFD conditions (n=4-10, Mean±SEM, Unpaired t-test, *P < 0.05, ***P < 0.001).

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 13. Oil Red O staining of aortic valve in male Ldlr-KO mice and WT mice under HFD conditions. 

The results indicate that no lipid accumulation was observed in the aortic valves of male WT mice at 6 and 17 weeks of HFD, while all male Ldlr-KO mice (2/2) exhibited significant lipid accumulation in the aortic valves as early as 6 weeks of HFD, with the severity of this phenotype progressively worsened with age (n=2).

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 14. Oil Red O staining of aortic valve in female Ldlr-KO mice and WT mice under HFD conditions. 

The results indicate that no lipid accumulation was observed in the aortic valves of female WT mice at 6 and 17 weeks of HFD, while all female Ldlr-KO mice (2/2) exhibited significant lipid accumulation in the aortic valves as early as 6 weeks of HFD, with the severity of this phenotype progressively worsened with age (n=2).

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 15. Oil Red O staining of aorta in male Ldlr-KO mice and WT mice under HFD conditions (n=2). 

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 16. Oil Red O staining of aorta in female Ldlr-KO mice and WT mice under HFD conditions (n=2). 

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 17. Masson staining of aortic valve in male Ldlr-KO mice and WT mice under HFD conditions. 

Muscle fibers appear red, while collagen fibers are stained blue. Ldlr-KO mice exhibited increased collagen content, suggesting enhanced plaque stability (n=2). 

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 18. Masson staining of aortic valve in female Ldlr-KO mice and WT mice under HFD conditions. 

Muscle fibers appear red, while collagen fibers are stained blue. Ldlr-KO mice exhibited increased collagen content, suggesting enhanced plaque stability (n=2). 

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 19. Representative pictures of aortic valve. 

In male WT mice after 17 weeks of HFD feeding, the vascular intima of cardiac tissue appeared smooth with no plaque formation. Myocardial cells were loosely arranged (yellow polygon). However, male Ldlr-KO mice after 17 week of HFD feeding displayed abundant infiltration of foam cells were observed (yellow arrows), with a small amount of hyperplasia of fibrous connective tissue (orange arrows). Myocardial cells displayed loose and irregular arrangement. The black box indicates the magnified region.

Abbr. WT, wild type; HO, homozygous; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.

Fig 20. Representative pictures of aortic valve. 

In female WT mice after 17 weeks of HFD feeding, the vascular intima of cardiac tissue appeared smooth with no plaque formation. Myocardial cells were loosely arranged and brownish-yellow pigment deposition was observed on the valve (brown arrow). However, female Ldlr-KO mice after 17 week of HFD feeding displayed fibrous connective tissue (orange arrows) and a small amount of foam cell accumulation (yellow arrows) on the surface. A small amount of necrotic cell debris and unstructured necrotic material could be observed (purple arrows). A small amount of brownish-yellow pigment deposition was observed on the valve (brown arrow). Myocardial cells displayed loose and irregular arrangement. The black box indicates the magnified region.

Abbr. WT, wild type; HFD, high-fat diet.

Note. HFD started at 6 weeks of age.