Gene targeting strategy for B-hEPCAM MC38 cells. The exogenous promoter and human EPCAM coding sequence was inserted to replace part of murine exon 4 and all of exons 5~7. The insertion disrupts the endogenous murine Epcam gene, resulting in a non-functional transcript.

EPCAM expression analysis in B-hEPCAM MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hEPCAM MC38 cultures were stained with species-specific anti-EPCAM antibody. Human EPCAM was detected on the surface of B-hEPCAM MC38 cells but not wild-type MC38 cells. The 3-C12 clone of B-hEPCAM MC38 cells was used for in vivo experiments.