Expression of a putative ligand for CD112R. (A) Immune cells in human blood and monocyte-derived DCs were stained with control (FLAG-Fc; red) or CD112R-Fc (blue) protein by flow cytometry. Immune cells were gated on individual lineage markers. T cell: CD3; B cell: CD19; NK cell: CD56⁺CD3⁻; monocyte: CD14; DC: CD11c. (B) Cultured human tumor cell lines were stained with control (FLAG-Fc) and CD112R protein, and the median fluorescence intensity (MFI) was determined by flow cytometry. CD112R MFI ratios (CD112R MFI/control MFI) are indicated. (C) SK-MEL28 melanoma line with or without trypsin treatment (10 min) was stained with control (FLAG-Fc; red) or CD112R-Fc (blue) protein by flow cytometry. (D) Blockade of CD112R binding by a CD112R mAb. HEK293T cells were stained with control (FLAG-Fc; red) or CD112R-Fc (blue) protein by flow cytometry with or without the presence of a CD112R mAb (clone 2H6). All data shown are representative of at least three independent experiments.