Basic Information
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Targeting strategy
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The exogenous promoter and human B7-H3 coding sequence were inserted to replace part of murine exon 3 and all of exon 4. The insertion disrupts the endogenous murine B7h3 gene, resulting in a non-functional transcript.
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Protein expression analysis
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B7-H3 expression analysis in B-hB7-H3 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hB7-H3 MC38 cultures were stained with species-specific anti-B7-H3 antibody. Human B7-H3 was detected on the surface of B-hB7-H3 MC38 cells but not wild-type MC38 cells. The 2-F02 clone of B-hB7-H3 MC38 cells was used for in vivo experiments.
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Tumor growth curve
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Subcutaneous homograft tumor growth of B-hB7-H3 MC38 cells. B-hB7-H3 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N mice (female, 5-8-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Bodyweight (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-hB7-H3 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Recommendations for inoculation in other mouse strains:
1. Cell inoculation amount is recommended to be tried between 5×105-1×107;
2.Inoculated cells can be tried to be suspended with DMEM stock solution.