Fig. 1

ZG16 binds to glycosylated PD-L1 through its lectin domain a Structure of ZG16 and PD-L1. b-c Co-immunoprecipitation of Flag-tagged ZG16 (ZG16-Flag) with His-tagged PD-L1 (PD-L1-His). Plasmid construct of ZG16-Flag was co-transfected with PD-L1-His into SW480 cells. Single vectors expressing each tag (Flag, His) were used as negative controls. d Immunofluorescence of SW480 cells transfected with ZG16-Flag and PD-L1-His, alone or in combination, for 24 h. Cells were subsequently stained with antibodies against His-tag (Red) and Flag-tag (Green) and DAPI (blue; nuclei). e Structure of ZG16-D151A, ZG16-M5, and PD-L1-4NQ f Co-immunoprecipitation of Flag-tagged ZG16 (ZG16-Flag, ZG16-D151A-Flag or ZG16-M5-Flag) with His-tagged PD-L1 (PD-L1-His). Plasmid constructs of ZG16-Flag, ZG16-D151A-Flag, or ZG16-M5-Flag were co-transfected with PD-L1-His into SW480 cells. Single vectors expressing each tag (Flag, His) were used as negative controls. g Co-immunoprecipitation of His-tagged PD-L1 (PDL1-His or PDL1-4NQ-His) with Flag-tagged ZG16 (ZG16-Flag). Plasmid constructs of PDL1-His or PDL1-4NQ-His were co-transfected with ZG16-Flag into SW480 or HCT118 cells. Single vectors expressing each tag (Flag, His) were used as negative controls. h Immunoblots of CD3⁺T cells cocultured withSW480 or SW480-ZG16 for 24 h. i INF-r expression of CD4⁺T cells or CD8⁺T cells cocultured withSW480 or SW480-ZG16. j Jurkat cells were treated with purified ZG16 protein for 48 h and then western blot was performed to detect IFNr expression