hIL1R1

利用同源重组，将小鼠Il1r1基因进行人源化修饰。

Fig.1 Detection of IL1R1 expression in spleen by RT-PCR. Mouse Il1r1 mRNA (258 bp) was detectable only in wild-type C57BL/6 mice. Human IL1R1 mRNA (227 bp) was detectable only in homozygous hIL1R1 knockin mice but not in wild-type mice. Spleen RNA was extracted from 10-week-old male wild-type C57BL/6 (WT)(n=1) and homozygous hIL1R1 knockin mice (HO) (n=2), 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 IL1R1 expression in bladder and spleen by RT-PCR. Mouse Il1r1 mRNA (258 bp) was detectable only in wild-type C57BL/6 mice. Human IL1R1 mRNA (227 bp) was detectable only in homozygous hIL1R1 knockin mice but not in wild-type mice. Bladder and spleen RNA was extracted from 6-week-old female wild-type C57BL/6 (WT)(n=1) and homozygous hIL1R1 knockin mice (HO) (n=2), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

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

Fig.3 Detection of IL1R1 expression in hIL1R1 mice by WB. IL1R1 was detectable in spleen from both WT C57BL/6 and HO hIL1R1 mice , as the antibody was cross-reactived between human and mouse IL1R1. The spleen tissue lysates were collected from 10-week-old male wild-type C57BL/6 mice and 10-week-old male homozygous hIL1R1 mice, and then analyzed by western blot with anti-IL1R1 antibody. 

Abbr. HO, homozygous; WT, wild type; PC, positive control, Hela cells.

Fig.4 Detection of mouse IL-6 expression in serum of WT C57BL/6 mice and HO hIL1R1 mice by ELISA. mIL-6 was detectable in serum from both WT C57BL/6 and HO hIL1R1 mice. Serum were collected from wild-type C57BL/6 mice (n=2, female, 10 weeks old) and HO hIL1R1 mice (n=2, female, 10 weeks old), and analyzed by ELISA with anti-mIL6 ELISA kit. 

Abbr.  HO, homozygous; WT, wild type.

Note. hIL1R1 and C57BL/6 mice were i.p. injected with IL-1β(1μg/mouse) for 2h.