The exogenous promoter and mouse Efna3 coding sequence were inserted into the mouse genome randomly. Mouse Efna3 is highly expressed on the surface of B-Tg(mEfna3) MC38 cells.
Application
B-Tg(mEfna3) MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.
Targeting strategy
Gene targeting strategy for B-Tg(mEfna3) MC38 cells. The exogenous promoter and mouse Efna3 coding sequence were inserted into the mouse genome randomly.
Protein expression analysis
Efna3 expression analysis in B-Tg(mEfna3) MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-Tg(mEfna3) MC38 cultures were stained with species-specific anti-Efna3 antibody. Mouse Efna3 was detected on the surface of B-Tg(mEfna3) MC38 cells but not wild-type MC38 cells. The 2-G06 clone of B-Tg(mEfna3) MC38 cells was used for in vivo experiments.
Tumor growth curve & Body weight changes
Subcutaneous homograft tumor growth of B-Tg(mEfna3) MC38 cells. B-Tg(mEfna3) MC38 cells (5x105) and wild-type MC38 cells (5x105) were subcutaneously implanted into C57BL/6 mice (female, 5-week-old, n=6). 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-Tg(mEfna3) MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Tumor growth curve of individual mice
B-Tg(mEfna3) MC38 tumor cells growth of individual mice. B-Tg(mEfna3) MC38 cells (5x105) and wild-type MC38 cells (5x105) were subcutaneously implanted into C57BL/6 mice (female, 5-week-old, n=6). As shown in panel, B-Tg(mEfna3) MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Protein expression analysis of tumor cells
B-Tg(mEfna3) MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6), and on 25 days post inoculation, tumor cells were harvested and assessed for human Efna3 expression by flow cytometry. As shown, human Efna3 was highly expressed on the surface of tumor cells. Therefore, B-Tg(mEfna3) MC38 cells can be used for in vivo efficacy studies of novel Efna3 therapeutics.