Basic Information
Description
The mouse Plaur gene was replaced by human PLAUR coding sequence in B-hPLAUR MC38 cells. Human PLAUR is highly expressed on the surface of B-hPLAUR MC38 cells.
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Targeting strategy
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Gene targeting strategy for B-hPLAUR MC38 cells. The exogenous promoter and chimeric CDS containing mouse signal peptide and human PLAUR coding sequence was inserted to replace part of mouse Plaur. The insertion disrupts the endogenous murine Plaur gene, resulting in a non-functional transcript.
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Protein expression analysis
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PLAUR expression analysis in B-hPLAUR MC38 cells by flow cytometry. Single cell suspensions from B-hPLAUR MC38 cultures were stained with species-specific anti-PLAUR antibody. Human PLAUR was detected on the surface of B-hPLAUR MC38 cells but not wild-type MC38 cells. The 5-F05 clone of B-hPLAUR 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-hPLAUR MC38 cells. B-hPLAUR MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into B-hPLAUR mice (male, 7-9-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-hPLAUR MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Tumor volume and weight measurements
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Protein expression analysis of tumor cells
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B-hPLAUR MC38 cells were subcutaneously transplanted into B-hPLAUR mice (n=5), and on 35 days post inoculation, tumor cells were harvested and assessed for human PLAUR expression by flow cytometry. As shown, human PLAUR was highly expressed on the surface of tumor cells. Therefore, B-hPLAUR MC38 cells can be used for in vivo efficacy studies of novel PLAUR therapeutics.
