Description
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.
Targeting strategy
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.
Protein expression analysis
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.
Protein expression analysis of tumor cells
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.
Tumor growth curve & Body weight changes
Subcutaneous homograft tumor growth of B-hAMHR2 MC38 cells. B-hAMHR2 MC38 cells (5x105) and wild-type MC38 cells (5x105) were subcutaneously implanted into C57BL/6N mice (female, 7-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hAMHR2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Subcutaneous homograft tumor growth of B-hAMHR2 MC38 cells. B-hAMHR2 MC38 cells and wild-type MC38 cells were subcutaneously implanted into B-h4-1BB mice (female, 6-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hAMHR2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.
