Gene targeting strategy for B-CAG-hHER2 EL4.The exons 2-7 of mouse Her2 gene that encode the extracellular domain were replaced by human HER2 CDS region in B-CAG-hHER2 EL4.
Protein expression analysis
The HER2 expression analysis in B-CAG-hHER2 EL4 cells by flow cytometry. Single cells were collected from wild-type EL4 and B-CAG-hHER2 EL4 cells, and analyzed by flow cytometry with species-specific anti-HER2 antibody. Human HER2 was detectable in B-CAG-hHER2 EL4 cells but not wild-type EL4 cells.
Tumor Growth Curve & Body Weight Changes
Subcutaneous xenograft tumor growth of B-CAG-hHER2 EL4 cells. B-CAG-hHER2 EL4 cells and wild-type MC38 cells(2×105) were subcutaneously implanted into the C57BL/6N mice (female, 6-9 week-old, n=5).Tumor size and mice body weight were measured twice a week. (A) Tumor average volume ± SEM, (B) Mice body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5a X b2, where a and b were the long and short diameters of the tumor, respectively. As shown in panel A, B-CAG-hHER2 EL4 were able to establish tumor in vivo and can be used for efficacy study.
Tumor expression detection sample information
Protein expression analysis in tumor cells
FMO Control 1: B-CAG-hHER2 EL4 tumor cells only stained anti-mHERT2 PE antibody;
FMO Control 2: B-CAG-hHER2 EL4 tumor cells only stained anti-hHERT2 AF647 antibody
B-CAG-hHER2 EL4 cells were transplanted into C57BL/6N mice, and on 32 days post inoculation, certain volumes of tumors as was taken to detect the expression level of the cells. As Shown,hHER2 was highly expressed on the surface of tumor cells. B-CAG-hHER2 EL4 can be used for in vivo efficacy validation.