hCXCR4(2)

Fig.1 Detection of CXCR4 expression in thymus by RT-PCR. Mouse Cxcr4 mRNA (248 bp) was detectable only in wild-type C57BL/6 mice. Human CXCR4 mRNA (319 bp) was detectable only in homozygous hCXCR4(2) knockin mice but not in wild-type mice. Thymus RNA was extracted from 6-week-old female wild-type C57BL/6 (WT)(n=2) and homozygous hCXCR4(2) knockin mice (HO) (n=2), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. 

Fig.2 Detection of CXCR4 expression on T, B cells and CD11b+ cells in spleen of WT C57BL/6 mice and HO hCXCR4(2) mice by FACS. hCXCR4 was exclusively detectable in hCXCR4(2) humanized mice but not in wild-type mice. hCXCR4 was exclusively detectable on T, B cells and CD11b+ cells in hCXCR4(2) humanized mice, whereas mCXCR4 was exclusively detectable on T, B cells and CD11b+ cells in wild-type C57BL/6 mice. Leukocytes were collected from spleen of wild-type C57BL/6 mice (female, 5 weeks old) and homozygous hCXCR4(2) humanized mice (female, 6 weeks old), and analyzed by flow cytometry with anti-mCXCR4 and anti-hCXCR4. 

Abbr. HO, homozygous; WT, wild type.

Fig.3 Detection of CXCR4 expression on T, B cells and CD11b+ cells in BM of WT C57BL/6 mice and HO hCXCR4(2) mice by FACS. hCXCR4 was exclusively detectable in hCXCR4(2) humanized mice but not in wild-type mice. hCXCR4 was exclusively detectable on T, B cells and CD11b+ cells in hCXCR4(2) humanized mice, whereas mCXCR4 was exclusively detectable on T, B cells and CD11b+ cells in wild-type C57BL/6 mice. Leukocytes were collected from BM of wild-type C57BL/6 mice (female, 5 weeks old) and homozygous hCXCR4(2) humanized mice (female, 6 weeks old), and analyzed by flow cytometry with anti-mCXCR4 and anti-hCXCR4. 

Abbr. HO, homozygous; WT, wild type.

Fig.4 Detection of ulocuplumab+ on T, B cells and CD11b+ cells in BM of WT C57BL/6 mice and HO hCXCR4(2) mice by FACS. Ulocuplumab+ cells was exclusively detectable in hCXCR4(2) humanized mice but not in wild-type mice.  Leukocytes were collected from BM of wild-type C57BL/6 mice (female, 5 weeks old) and homozygous hCXCR4(2) humanized mice (female, 6 weeks old), and analyzed by flow cytometry with ulocuplumab and anti-human IgG. 

Abbr. HO, homozygous; WT, wild type.

Note. The positive signals observed in wild-type mice were due to non-specific staining, with a positive rate comparable to that of the FMO (fluorescence minus one) control.