hAPOE4/5xFAD

Fig.1 Detection of APOE  expression in liver by RT-PCR. Mouse Apoe mRNA (301 bp) was detectable only in wild-type C57BL/6 mice. Human APOE mRNA (139 bp) was detectable only in homozygous hAPOE4/5×FAD knockin mice but not in wild-type mice. Liver RNA was extracted from 14-week-old and 22-week-old  male wild-type C57BL/6 (WT)(n=6) and homozygous hAPOE4/5×FAD knockin mice (HO) (n=6), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.2 Detection of human APOE expression in liver of WT C57BL/6 mice and HO/HE hAPOE4/5xFAD mice by ELISA.  APOE was detectable in the liver from WT C57BL/6 mice and HO/HE hAPOE4/5xFAD mice, Human APOE was exclusively detectable in hAPOE4/5xFAD knockin mice，as the ELISA kit was cross-reactived between human and mouse. Liver tissues lysates were collected from wild-type C57BL/6 mice (n=3, male) and HO/HE hAPOE4/5xFAD mice (n=3, male), and analyzed by ELISA with anti-hAPOE ELISA kit. 

Abbr.  HO, homozygous; HE, heterozygote; WT, wild type.

Fig.3 Detection of human APOE expression in hippocampus of WT C57BL/6 mice and HO/HE hAPOE4/5xFAD mice by ELISA.  APOE was detectable in the hippocampus from WT C57BL/6 mice and HO/HE hAPOE4/5xFAD mice, Human APOE was exclusively detectable in hAPOE4/5xFAD knockin mice，as the ELISA kit was cross-reactived between human and mouse. Hippocampus tissues lysates were collected from wild-type C57BL/6 mice (n=3, male) and HO/HE hAPOE4/5xFAD mice (n=3, male), and analyzed by ELISA with anti-hAPOE ELISA kit. 

Abbr.  HO, homozygous; HE, heterozygote; WT, wild type.

Fig.4 Detection of human APOE expression in cortex of WT C57BL/6 mice and HO/HE hAPOE4/5xFAD mice by ELISA.  APOE was detectable in the cortex from WT C57BL/6 mice and HO/HE hAPOE4/5xFAD mice, Human APOE was exclusively detectable in hAPOE4/5xFAD knockin mice，as the ELISA kit was cross-reactived between human and mouse. Cortex tissues lysates were collected from wild-type C57BL/6 mice (n=3, male) and HO/HE hAPOE4/5xFAD mice (n=3, male), and analyzed by ELISA with anti-hAPOE ELISA kit. 

Abbr.  HO, homozygous; HE, heterozygote; WT, wild type.

Fig.5 Detection of human APOE expression in the hippocampus and cortex of hAPOE4/5xFAD mice by IHC. Compared with WT C57BL/6 mice, APOE expression was significantly increased in the hippocampus and cortex of hAPOE4/5xFAD mice, with the difference becoming more pronounced with increasing age. Brain tissues were harvested from mice (n=2 males), fixed with 4% paraformaldehyde, cryosectioned in the coronal plane, and stained by immunohistochemistry.