Figure 3.

Activated Rab8 missorts only AP-1B cargo. (A–C) Fully polarized MDCK cells were microinjected with the cDNAs encoding activated GFP-Rab8 (Rab8Q67L; A, second column), nonprenylated GFP-Rab8 (Rab8ΔC; A, third column), activated GFP-Rab11 (Rab11Q70L; A, fourth column), or dominant-negative GFP-Cdc42 (Cdc42T17N) (C) with LDLR (A and C, red) or FcR (B and C, red). After injection, the cells were incubated at 37°C for 1 h, at 20°C for 2.5 h, and finally at 37°C for 2 h in the presence of cycloheximide. Cells were fixed without permeabilization and stained for surface LDLR (C7) or FcR (24G2), followed with Alexa^® 568 secondary antibodies. GTPases are visualized by GFP fluorescence. Images are representative confocal z-sections.