Figure 6.

Activated Rab8 disrupts γ-adaptin localization in MDCK cells, but does not disrupt Golgi- or furin-containing regions of the TGN. (A) Top: MDCK cells grown on coverslips were microinjected with the cDNA for T7- Rab8Q67L (200 ng/μl), incubated at 37°C for 2 h, fixed, permeabilized, and stained with anti-γ-adaptin antibody 100/3 (red, second column) or anti-T7 (green, third column). Arrows, cells expressing Rab8Q67L where γ-adaptin localization is disrupted; arrowheads, cells not expressing Rab8 with normal γ-adaptin localization. Bottom: cells were injected with cDNA for nonprenylated, activated GFP-Rab8ΔC (green, third column). Arrowheads, cells expressing Rab8ΔC have normal γ-adaptin localization (red, second column). (B) MDCK cells were microinjected with cDNA of T7-Rab8Q67L, incubated at 37°C for 2 h, and processed for IF with the TGN antibody anti-furin (A; top, second column), for the Golgi with anti-giantin (B; bottom, second column), and for T7-Rab8 (A and B, third column).