Figure 3
From: Development of anti-feline PD-1 antibody and its functional analysis
Inhibition of fPD-1 and fPD-L1 interaction by mAbs. (A) Western blotting of soluble fPD1-hIg and fPDL1-hIg fusion proteins. fPD1-hIg, fPDL1-hIg, and human IgG2-Fc (hIg) were purified from the supernatant of Expi-293F cells and submitted to western blotting with anti-human IgG-horseradish peroxidase (HRP). Full image of western blotting is shown in Supplementary Figure for Fig. 3A. (B) NIH3T3 or mock and NIH3T3/fPD1 and NIH3T3/fPD1 were stained with dose-titrated fPDL1-hIg fusion protein (left panel), or NIH3T3 or mock and NIH3T3/fPDL1 and NIH3T3/fPDL1 with fPD1-hIg (right panel) and followed by staining with a secondary anti-human IgG-PE antibody. (C) NIH3T3/fPD1 cells were preincubated with serially titrated anti-PD-1 (1A1-2) mAb, and, after washing, incubated with 1 μg/mL of fPDL1-hIg or human Ig (left panel). NIH3T3/fPDL1 cells were preincubated with anti-PD-L1 (G11-6) mAb, and, after washing, incubated with 5 μg/mL of fPD1-hIg or human Ig (right panel). Cells were then stained with a secondary antibody anti-human IgG-PE.
