FIGURE 2.

Inhibition of PD‐L1 decreases phosphorylation of MET Tyr1234/5. A‐F, Cells were transfected with various concentrations of PD‐L1‐siRNA or control siRNA for 48 h, and western blot was used to analyze the knockdown efficiency and changes in MET phosphorylation in A375 (A, B), A431 (C, D), and HCC827 (E, F). G, Effect of PD‐L1 inhibition on HGF‐induced MET phosphorylation. Cells were treated with 40 μg/mL HGF for 30 min after the cells were transfected with PD‐L1 siRNA, and the changes in MET phosphorylation were analyzed by western blot. H, Effects of PD‐L1 inhibition by PD‐L1 antibody on MET activation. Cells were treated with 40 μg/mL PD‐L1 antibody for 48 h, and changes in MET phosphorylation were analyzed by western blot. I, Effects of PD‐L1 inhibition by PD‐1 recombinant protein on MET activation. Cells were treated with 40‐1000 ng/mL PD‐1 recombinant protein for 36 h, and the changes in MET phosphorylation were analyzed by western blot. J, Effects of PD‐L1 inhibition by PD‐1 recombinant protein on MET activation. Cells were treated with 1000 ng/mL PD‐1 recombinant protein for 12, 24, and 36 h, and changes in MET phosphorylation were analyzed by western blot. Bands were quantified using ImageJ software and quantitative data are presented as mean ± SD based on 3 biological replicates