Apoe-KO(2)/hAPOC3

1. Chow diet conditions：

Fig.1 hAPOC3 levels in Apoe-KO(2)/hAPOC3 mice under CD conditions. Under CD conditions, hAPOC3 was detectable in homozygous Apoe-KO(2)/hAPOC3 mice. Serum hAPOC3 levels in Apoe-KO(2)/hAPOC3 mice slightly decreased with age. Serum samples were collected following a 6-hour fast at different ages to measure levels of serum hAPOC3 (n=4-12, Mean ± SEM).

 Abbr. WT, wild-type; HO, homozygous; WD, Western diet.

Fig.2 Body weight change of Apoe-KO(2)/hAPOC3 mice under CD conditions. Apoe-KO(2)/hAPOC3 mice exhibited comparable body weight gain to both Apoe-KO(2) and hAPOC3 mice within the same sex. WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice were maintained on CD feeding. Body weight and body weight gain were recorded weekly (n=4-12, Mean ± SEM). 

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

Fig.3 Blood lipid profiles of Apoe-KO(2)/hAPOC3 mice after ARO-APOC3 treatment under CD conditions. Male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice were maintained on CD feeding. Serum samples were collected following a 6-hour fast at different ages to measure levels of (A) triglycerides (TG), (B) total cholesterol (T-CHO), (C) high-density lipoprotein cholesterol (HDL-C) and (D) low-density lipoprotein cholesterol (LDL-C) (n=4-12, Mean ± SEM, t-test, Apoe-KO(2) mice versus Apoe-KO(2)/hAPOC3 mice: #p<0.05, ##p<0.01; hAPOC3 mice versus Apoe-KO(2)/hAPOC mice: **p<0.01, ***p<0.001, ****p<0.0001). 

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

Fig.4 Blood lipid profiles of Apoe-KO(2)/hAPOC3 mice after ARO-APOC3 treatment under CD conditions. Under CD conditions, female Apoe-KO(2)/hAPOC3 mice exhibited significantly elevated levels of TG, T-CHO, and LDL-C, along with decreased levels of HDL-C, compared to hAPOC3 mice. Female WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice were maintained on CD feeding. Serum samples were collected following a 6-hour fast at different ages to measure levels of (A) triglycerides (TG), (B) total cholesterol (T-CHO), (C) high-density lipoprotein cholesterol (HDL-C) and (D) low-density lipoprotein cholesterol (LDL-C) (n=5-12, Mean ± SEM, t-test, Apoe-KO(2) mice versus Apoe-KO(2)/hAPOC3 mice: #p<0.05, ##p<0.01; hAPOC3 mice versus Apoe-KO(2)/hAPOC mice: **p<0.01, ***p<0.001, ****p<0.0001). 

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

Fig.5 Oil Red O staining of aortas of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice under CD conditions (n=3 in each group, 3 months old). No significant lipid accumulation was observed in aortas in WT mice, Apoe-KO(2) mice and hAPOC3 mice at 3 months of age under chow diet conditions. In contrast, Apoe-KO(2)/hAPOC3 mice exhibited slight lipid accumulation near the aortic arch at 3 months of age under chow diet conditions.

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

Fig.6 Oil Red O staining of aortas of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice under CD conditions (n=3 in each group, 5 months old). No significant lipid accumulation was observed in the aortas of WT mice and hAPOC3 mice at 5 months of age under chow diet conditions. In contrast, slight lipid deposition was detected in Apoe-KO(2) mice, whereas Apoe-KO(2)/hAPOC3 mice showed significant lipid accumulation near the aortic arch at 5 months of age under chow diet conditions.

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

Fig.7 Oil Red O staining of aortas of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice under CD conditions (n=3 or 4 in each group, 7 months old). No significant lipid accumulation was observed in the aortas of WT mice and hAPOC3 mice at 7 months of age under chow diet conditions. In contrast, Apoe-KO(2) mice and Apoe-KO(2)/hAPOC3 mice showed significant lipid accumulation near the aortic arch at 7 months of age under chow diet conditions.

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

Fig.8 Oil Red O staining of aortic valves of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice at 3, 5 and 7 months of age under CD conditions (n=3 or 4 in each group). The sacle bar is 0.5 mm. No lipid accumulation was observed in the aortic valves of WT or hAPOC3 mice at any age. In contrast, both Apoe-KO(2) and Apoe-KO(2)/hAPOC3 mice showed slight lipid accumulation in the aortic valves at 3 months of age, with a significant increase observed at 5 and 7 months.

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

Fig.9 Oil Red O staining of aortas of female WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice under CD conditions (n=3 in each group, 3 months old). No significant lipid accumulation was observed in aortas in all mice at 3 months of age under chow diet conditions. 

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

Fig.10 Oil Red O staining of aortas of female WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice under CD conditions (n=3 in each group, 5 months old). No significant lipid accumulation was observed in the aortas of WT mice and hAPOC3 mice at 5 months of age under chow diet conditions. In contrast, slight lipid deposition was detected in Apoe-KO(2) mice and Apoe-KO(2)/hAPOC3 mice near the aortic arch at 5 months of age under chow diet conditions.

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

Fig.11 Oil Red O staining of aortas of female WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice under CD conditions (n=3 in each group, 7 months old). No significant lipid accumulation was observed in the aortas of hAPOC3 mice at 7 months of age under chow diet conditions. In contrast, Apoe-KO(2) mice and Apoe-KO(2)/hAPOC3 mice showed significant lipid accumulation near the aortic arch at 7 months of age under chow diet conditions.

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

Fig.12 Oil Red O staining of aortic valves of female WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice at 3, 5 and 7 months of age under CD conditions (n=2 or 3 in each group). No lipid accumulation was observed in the aortic valves of WT or hAPOC3 mice at any age. In contrast, both Apoe-KO(2) and Apoe-KO(2)/hAPOC3 mice showed slight lipid accumulation in the aortic valves at 3 months of age, with a significant increase observed at 5 and 7 months.

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

2. Western diet conditions：

Fig.13 hAPOC3 levels in male Apoe-KO(2)/hAPOC3 mice under WD conditions. Under WD conditions, hAPOC3 was detectable in homozygous Apoe-KO(2)/hAPOC3 mice. Serum hAPOC3 levels in Apoe-KO(2)/hAPOC3 mice were comparable across 8 weeks to 18 weeks of age. Serum samples were collected following a 6-hour fast at different ages to measure levels of serum hAPOC3 (n=4-6, Mean ± SEM).

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.

Fig.14 Blood lipid profiles of Apoe-KO(2)/hAPOC3 mice after ARO-APOC3 treatment under WD conditions. Under WD conditions, male Apoe-KO(2)/hAPOC3 mice exhibited significantly elevated levels of TG, T-CHO, and LDL-C, along with decreased levels of HDL-C, compared to hAPOC3 mice. Male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice were maintained on WD feeding from 6 weeks of age. Serum samples were collected following a 6-hour fast at different ages to measure levels of (A) triglycerides (TG), (B) total cholesterol (T-CHO), (C) high-density lipoprotein cholesterol (HDL-C) and (D) low-density lipoprotein cholesterol (LDL-C) (n=4-6, Mean ± SEM, t-test, Apoe-KO(2) mice versus Apoe-KO(2)/hAPOC3 mice: #p<0.05; hAPOC3 mice versus Apoe-KO(2)/hAPOC mice: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). 

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.

Fig.15 Oil Red O staining of aortas of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice after 7 weeks of WD feeding (n=2 in each group, 13 weeks old). No significant lipid accumulation was observed in the aortas of all mice after 7 weeks of WD feeding. WD feeding was start at 6 weeks of age.

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.

Fig.16 Oil Red O staining of aortas of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice after 12 weeks of WD feeding (n=2 in each group, 18 weeks old). After 12 weeks of WD feeding, no significant lipid accumulation was observed in the aortas of WT mice and hAPOC3 mice. Conversely, Apoe-KO(2) mice and Apoe-KO(2)/hAPOC3 mice showed significant lipid accumulation near the aortic arch. WD feeding was start at 6 weeks of age.

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.

Fig.17 Oil Red O staining of aortic valves of male WT C57BL/6 mice, HO Apoe-KO(2) mice, HO hAPOC3 mice and HO/HO Apoe-KO(2)/hAPOC3 mice after 7 weeks or 12 weeks of Western diet feeding (n=2 or 4 in each group). WD feeding was start at 6 weeks of age. No lipid accumulation was observed in the aortic valves of WT or hAPOC3 mice at any timepoint. In contrast, both Apoe-KO(2) and Apoe-KO(2)/hAPOC3 mice showed slight lipid accumulation in the aortic valves after 7 weeks of Western diet feeding, with a significant increase observed after 12 weeks of Western diet feeding.

Abbr. WT, wild-type; HO, homozygous; WD, Western diet.