Basic Information
Description
The mouse Anxa2 gene was replaced by human ANXA2 coding sequence in B-hANXA2 MC38 cells. Human ANXA2 is highly expressed on the surface of B-hANXA2 MC38 cells.
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Targeting strategy
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Gene targeting strategy for B-hANXA2 MC38 cells. The exogenous promoter and human ANXA2 coding sequence was inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Anxa2 gene, resulting in a non-functional transcript.
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Protein expression analysis
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ANXA2 expression analysis in B-hANXA2 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hANXA2 MC38 cultures were stained with anti-ANXA2 antibody. Human ANXA2 was detected on the surface of B-hANXA2 MC38 cells and MC38 cells, as the antibody is crossly reactive with ANXA2 in human and mice. It has been verified that B-hANXA2 MC38 cells do not express mouse mRNA by RT-PCR. The 2-A01 clone of B-hANXA2 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-hANXA2 MC38 cells. B-hANXA2 MC38 cells (1×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old). 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 diaANXA2er X short diaANXA2er2. As shown in panel A, B-hANXA2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.