Fig. 1

PTN97–110 inhibits the interaction of PTN and VEGFA with PTPRZ1. (A) Schematic representation of the domains that correspond to the studied PTN peptides on the tertiary structure of the PTN protein. The arrows represent the β-sheet domains, and the cylinders represent the random coils at the N- and C-terminal PTN domains. The five disulfide bonds on the PTN molecule are marked as grey. The numbers correspond to the respective amino acids of the secreted protein. The exact amino acid sequences of the PTN peptides tested in the present study are shown at the right. (B) C6 cells were cultured for 24 h in the presence of the tested peptides (100 ng/ml in the culture medium). Total cell lysates were then immunoprecipitated by using an antibody against PTPRZ1 or non-specific IgG and analyzed by Western blot using an antibody against PTN or VEGFA. The content of β-actin in the corresponding lysate was used as a loading control. Representative images from two independent experiments are shown. (C) HUVEC or LMVEC were incubated in the presence or absence of PTN97-110 (100 ng/ml) for 24 h. Total cell lysates were then immunoprecipitated by using an antibody against PTPRZ1 or non-specific IgG and analyzed by Western blot using an antibody against PTN or VEGFA. The content of β-actin in the corresponding lysate was used as a loading control. Representative images from two independent experiments are shown.