FIG. 3.
G-S is retained mostly in the ERGIC, and mutation of the dilysine motif results in transport to the plasma membrane and endocytosis. (A) HeLa cells expressing G-S were fixed, permeabilized, stained with either rabbit or mouse anti-VSV-G, and double labeled with either rabbit anticalnexin, mouse anti-ERGIC-53, or mouse anti-GM130. For cells double labeled with calnexin, the secondary antibodies were fluorescein-conjugated donkey anti-rabbit IgG and Texas Red-conjugated goat anti-mouse IgG. For cells double labeled with ERGIC-53 or GM130, secondary antibodies were fluorescein-conjugated goat anti-mouse IgG and Texas Red-conjugated donkey rabbit IgG. (B) Intact HeLa cells expressing G-S2A and G-S4A were incubated with mouse anti-VSV-G at 37°C for 15 min, fixed, permeabilized, and then stained with rabbit anti-VSV-G for internal expression. Secondary antibodies were fluorescein-conjugated donkey anti-rabbit IgG and Texas Red-conjugated goat anti-mouse IgG. Bars, 10 μm.