Characterization of anti-TIM-2 mAbs. (a) Reactivity of anti-T cell immunoglobulin and mucin domain (TIM)-2 monoclonal antibodies (mAbs) to mouse TIM family molecules. NRK-52E-derived TIM transfectants and parental cells were stained with biotinylated RMT2-14, RMT2-25, RMT2-26, specific mAbs against each TIM family molecule, or control IgG followed by PE-labeled streptavidin. Thick lines indicate the staining with the respective mAb and the dotted lines indicate background staining with control IgG. (b) H-ferritin binds to TIM-2 transfectant. NRK-52E-derived TIM transfectants were stained with Alexa647-labeled H-ferritin. Thick lines indicate the staining with the H-ferritin and the dotted lines indicate background staining with PBS. (c) Anti-TIM-2 mAbs inhibit H-ferritin binding to TIM-2 transfectant. TIM-2/L5178Y cells were pretreated with the indicated anti-TIM-2 mAb (thick lines) or control IgG (solid lines) and then stained with Alexa647-labeled H-ferritin. The dotted lines indicate background staining with PBS.