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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.