Gene targeting strategy for B-CAG-hCD47 MC38.The exons 2-3 of mouse CD47 gene that encode the extracellular domain were replaced by human CD47 CDS region in B-CAG-hCD47 MC38.
Protein expression analysis
The CD47 expression analysis in B-CAG-hCD47 MC38 cells by flow cytometry. Single cells were collected from wild-type MC38 and B-CAG-hCD47 MC38 cells, and analyzed by flow cytometry with species-specific anti-CD47 antibody. Human CD47 was detectable in B-CAG-hCD47 MC38 cells but not wild-type MC38 cells.
Tumor Growth Curve & Body Weight Changes
Subcutaneous xenograft tumor growth of B-CAG-hCD47 MC38 cells. B-CAG-hCD47 MC38 cells and wild-type MC38 cells (5×105) were subcutaneously implanted into the C57BL/6 mice (female, 5-6 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-hCD47 MC38 were able to establish tumor in vivo and can be used for efficacy study.