Gene targeting strategy for B-hITGA5 mice. The exons 1-29 of mouse Itga5 gene that encode the extracellular domain were replaced by human ITGA5 exons 1-29 in B-hITGA5 mice.

Strain specific analysis of ITGA5 gene expression in wild-type mice and B-hITGA5 mice by RT-PCR. Mouse Itga5 mRNA was detectable in bladder of wild-type mice (+/+). Human ITGA5 mRNA was detectable only in homozygous B-hITGA5 but not in wild-type mice.

Strain specific ITGA5 expression analysis in homozygous B-hITGA5 mice by flow cytometry. Splenocytes were collected from wild-type mice (+/+) and homozygous B-hITGA5 mice (H/H) stimulated with anti-CD3e in vivo (7.5 μg/mice, stimulation for 24 hours, i.p. ), and analyzed by flow cytometry with species-specific anti-ITGA5 antibody. Mouse ITGA5 was exclusively detectable in WT mice. Human ITGA5 were exclusively detectable in homozygous B-hITGA5 but not in wild-type mice.

Strain specific ITGA5 expression analysis in homozygous B-hITGA5 mice by flow cytometry. Splenocytes were collected from wild-type mice (+/+) and homozygous B-hITGA5 mice (H/H) stimulated with anti-CD3e in vivo (7.5 μg/mice, stimulation for 24 hours, i.p. ), and analyzed by flow cytometry with species-specific anti-ITGA5 antibody. Mouse ITGA5 was exclusively detectable in WT mice. Human ITGA5 were exclusively detectable in homozygous B-hITGA5 but not in wild-type mice.