Gene targeting strategy for B-CAG-hPD-L1 EL4. The human full-length CD274 CDS region was inserted in the Cd274 exon3 of wild-type EL4.
Protein expression analysis
The PD-L1 expression analysis in B-CAG-hPD-L1 EL4 cells by flow cytometry. Single cells were collected from wild-type EL4 and B-CAG-hPD-L1 EL4 cells, and analyzed by flow cytometry with species-specific anti-PD-L1 antibody. Mouse PD-L1 was detectable in wild-type EL4 cells. Human PD-L1 was exclusively detectable in B-CAG-hPD-L1 EL4 cells but not wild-type EL4 cells.
Tumor Growth Curve & Body Weight Changes
Subcutaneous xenograft tumor growth of B-CAG-hPD-L1 EL4 cells. B-CAG-hPD-L1 EL4 cells(2×105) were subcutaneously implanted into the C57BL/6 (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-hPD-L1 EL4 were able to establish tumor in vivo and can be used for efficacy study.