Figure 3.

Protein-protein docking for VISTA and VSIG3. (A) An overall map of the virtual docking of the human VISTA ECD (cyan) with the human VSIG3 ECD protein (green) based on the structures of hVISTA and hVSIG3. Key residues are depicted in stick representation (purple and orange). (B) Enlarged the schematic diagram of the binding site. (C) HEK293T cells were transfected with VSIG3-FLAG and VISTA-HA plasmids. Equal amounts of protein were immunoprecipitated with Anti-FLAG magnetic beads and immunoblotted with antibodies indicated. (D) Co-IP detected the interaction by multiple mutated amino acid epitope on VISTA and VSIG3. HEK293T cells were transfected with wild type (wt) or multiple point mutations (mul-mut) of VSIG3-FLAG or VISTA-HA plasmids. Equal amounts of protein were immunoprecipitated Anti-FLAG magnetic beads and immunoblotted with antibodies indicated.