Gene targeting strategy for B-hAMHR2 MC38 cells. The exogenous promoter and human AMHR2 coding sequence were inserted to replace part of murine exon 2 and all of exon 3. The insertion disrupts the endogenous murine Amhr2 gene, resulting in a non-functional transcript.

Gene targeting strategy for B-hAMHR2 MC38 cells. The exogenous promoter and human AMHR2 coding sequence was inserted to replace part of murine exon 2 and all of exon 3. The insertion disrupts the endogenous murine Amhr2 gene, resulting in a non-functional transcript.

AMHR2 expression analysis in B-hAMHR2 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hAMHR2 MC38 cultures were stained with species-specific anti-AMHR2 antibody. Human AMHR2 was detected on the surface of B-hAMHR2 MC38 cells, but not on the surface of wild-type MC38 cells. The 1-A01 clone of B-hAMHR2 MC38 cells was used for in vivo experiments.

B-hAMHR2 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human AMHR2 expression by flow cytometry. As shown, human AMHR2 was highly expressed on the surface of tumor cells. Therefore, B-hAMHR2 MC38 cells can be used for in vivo efficacy studies of novel AMHR2 therapeutics.