Fig. 3

Internalization and localization of GrB-Fc-4D5. a Internalization into SKOV3 target cells assessed by a polyclonal anti-GrB antibody demonstrated rapid internalization. SKOV3 cells were either untreated or treated with 50 nmol/L of GrB-Fc-4D5 for 1, 5.5 and 24 h. The cells were fixed, acid washed to remove surface bound fusion protein, and then permeabilized and immunostained for the presence of GrB by using a polyclonal rabbit anti-GrB antibody (green). Cells were counterstained with DAPI (blue) to identify nuclei and visualized using a fluorescent microscope. b Intracellular localization of GrB-Fc-4D5. A549 (top) and MDA-MB-231 (bottom) cells were treated with AF-594-GrB-Fc-4D5 (red) for two hours followed by addition of Lysotracker dye (blue). Cells were imaged live by confocal microscopy. Magnification, 20x