Basic Information
Description
The exogenous promoter and chimeric MERTK coding sequence containing the human signal peptide, extracellular domain, transmembrane domain and mouse cytoplasmic domain was inserted to replace part of murine exon 1.
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Targeting strategy
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Gene targeting strategy for B-hMERTK MC38 cells. The exogenous promoter and chimeric MERTK coding sequence containing human signal peptide, extracellular domain, transmembrane domain and mouse cytoplasmic domain was inserted to replace part of murine exon 1. The insertion disrupts the endogenous murine Mertk gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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MERTK expression analysis in B-hMERTK MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hMERTK MC38 cultures were stained with species-specific anti-MERTK antibody. Human MERTK was detected on the surface of B-hMERTK MC38 cells but not wild-type MC38 cells. The 1-G01 clone of B-hMERTK MC38 cells was used for in vivo experiments.
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Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-hMERTK MC38 cells. B-hMERTK MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 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-hMERTK MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
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Protein expression analysis of tumor cells
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B-hMERTK 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 MERTK expression by flow cytometry. As shown, human MERTK was highly expressed on the surface of tumor cells. Therefore, B-hMERTK MC38 cells can be used for in vivo efficacy studies of novel MERTK therapeutics.
