Gene targeting strategy for B-hGPC3 mice. The human GPC3 CDS was inserted after the exon 3 of mouse Gpc3 gene in B-hGPC3 mice. The insertion disrupts the endogenous murine Gpc3 gene.

Strain specific analysis of GPC3 gene expression in C57BL/6 wild-type (WT) mice and B-hGPC3 mice by RT-PCR. Mouse Gpc3 mRNA was detectable only in kidney of WT mice (+/+). Human GPC3 mRNA was detectable only in homozygous B-hGPC3 mice (H/H) but not in WT mice (+/+).

Immunohistochemical (IHC) analysis of GPC3 protein expression in C57BL/6 wild-type (WT) mice and B-hGPC3 mice. The liver and fetal liver were collected from C57BL/6 (+/+) and homozygous B-hGPC3 mice (H/H) and analyzed by IHC with anti-GPC3 antibody. Mouse GPC3 was detectable in WT mice, and human GPC3 was detectable in homozygous B-hGPC3 mice. The arrow indicates tissue cells with positive GPC3 staining (brown).