Fig. 8. Anti-LGP85 antibodies are internalized to early endosome-like large vacuoles. COS cells transfected with LGP85 for 36 hours were incubated for 1 hour with rabbit polyclonal antibodies against LGP85 at 4°C and chased for 3 hours at 37°C. Cells were then fixed, permeabilized, and incubated with a mouse monoclonal antibody to either EEA1 (A) or LAMP-2 (D). Localization of internalized and bound antibodies revealed by Alexa488-conjugated goat anti-rabbit IgG (green in B and E) and Alexa594-conjugated goat anti-mouse antibody (red in A and D), respectively, was visualized by confocal microscopy. The right columns (C,F) show the merged images of internalized antibodies (green in B and E) and EEA1 (red in A) or LAMP-2 (red in D). Bars, 20 µm.