hCRLF2/hIL7R/hTSLP

通过hCRLF2/hTSLP(NM-XA-241108)与hIL7R(NM-HU-210027)小鼠交配获得。

Fig.1 Detection of CRLF2 expression in adrenal gland by RT-PCR. Mouse Crlf2 mRNA (214 bp) was detectable only in WT C57BL/6 mice. Human CRLF2 mRNA (182 bp) was detectable only in HO/HO/HO hCRLF2/hIL7R/hTSLP mice but not in WT mice. Adrenal gland RNA was extracted from 9-week-old male WT C57BL/6 (WT)(n=3) and HO hCRLF2/hIL7R/hTSLP mice (HO/HO/HO) (n=3), 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 IL7R expression in spleen by RT-PCR. Mouse Il7r mRNA (310 bp) was detectable only in WT C57BL/6 mice. Human IL7R mRNA (167 bp) was detectable only in HO/HO/HO hCRLF2/hIL7R/hTSLP mice but not in WT mice. Spleen RNA was extracted from 9-week-old male WT C57BL/6 (WT)(n=3) and HO hCRLF2/hIL7R/hTSLP mice (HO/HO/HO) (n=3), 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 TSLP expression in bladder by RT-PCR. Mouse Tslp mRNA (160 bp) was detectable only in WT C57BL/6 mice. Human TSLP mRNA (168 bp) was detectable only in HO hCRLF2/hIL7R/hTSLP mice but not in WT mice. Bladder RNA was extracted from 9-week-old male WT C57BL/6 (WT)(n=3) and HO hCRLF2/hIL7R/hTSLP mice (HO/HO/HO) (n=3), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

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

Fig.4 Detection of IL7R expression on T cells in spleen of WT C57BL/6 mice and HO/HO/HO hCRLF2/hIL7R/hTSLP mice by FACS. hIL7R was exclusively detectable in hCRLF2/hIL7R/hTSLP humanized mice but not in WT mice，whereas mIL7R was exclusively detectable on T cells in WT C57BL/6 mice. Leukocytes were collected from spleen of wild-type C57BL/6 mice (male, 8 weeks old) and homozygous hCRLF2/hIL7R/hTSLP humanized mice (male, 8 weeks old) 4h post LPS stimulation（5mg/kg, i.p.) and analyzed by flow cytometry with anti-mIL7R and anti-hIL7R. 

Abbr. HO/HO/HO, homozygous; WT, wild type.

Fig.5 Detection of mTSLPR expression on DC and macrophage cells in spleen of WT C57BL/6 mice and HO/HO/HO hCRLF2/hIL7R/hTSLP mice by FACS. mTSLPR was exclusively detectable on DC and macrophage cells in wild-type mice but not in hCRLF2/hIL7R/hTSLP humanized mice. Leukocytes were collected from spleen of wild-type C57BL/6 mice (male, 8 weeks old) and homozygous hCRLF2/hIL7R/hTSLP humanized mice (male, 8 weeks old) 4h post LPS stimulation (5mg/kg, i.p.) and analyzed by flow cytometry with anti-mTSLPR. 

Abbr. HO/HO/HO, homozygous; WT, wild type.

Fig.6 Detection of hTSLPR expression on DC and macrophage cells in spleen of WT C57BL/6 mice and HO/HO/HO hCRLF2/hIL7R/hTSLP mice by FACS. hTSLPR was exclusively detectable on DC and macrophage cells in hCRLF2/hIL7R/hTSLP humanized mice but not in WT mice. Leukocytes were collected from spleen of wild-type C57BL/6 mice (male, 8 weeks old) and homozygous hCRLF2/hIL7R/hTSLP humanized mice (male, 8 weeks old) 4h-post LPS stimulation (5mg/kg, i.p.), and analyzed by flow cytometry with anti-mTSLPR. 

Abbr. HO/HO/HO, homozygous; WT, wild type.