Supplementary Figure 6: In situ zymography of frozen Ramos xenograft tumor slices.
From: A coiled-coil masking domain for selective activation of therapeutic antibodies
Protease activity was assessed by incubating Alexa Fluor 647-labeled hBU12 or CC3-IPVSLRSG-hBU12 antibodies (1 µg/mL) with fresh frozen Ramos tumor slices (5 µm). Proteolysis of the mask results in binding of the antibody, as detected using an Olympus fluorescence microscope. The addition of protease inhibitors resulted in reduced binding of CC3-IPVSLRSG-hBU12, whereas hBU12 binding was not impacted. Images were taken at 20x magnification and the scale bar indicates a 50 μm distance. Two independent experiments were conducted with similar results.
