FIGURE 4.

Inactive Rab13 traffics on vesicles following deletion of the C-terminal prenylation motif. A, schematic of the constructs used in the figure. FL, full-length; ΔC, C-terminal deletion of the last four amino acids CSLG (Cys-Ser-Leu-Gly); ΔHVD, hypervariable domain deletion. B, MCF10A cells expressing mCh-Rab13 Q67L FL, T22N FL, or T22N deletion constructs, as indicated, were imaged live. Scale bars = 10 μm. C, the percentage of transfected cells with mCh-Rab13 constructs on vesicles. Data are mean ± S.D., measuring a minimum of 20 cells/experiment from a minimum of three independent experiments. ***, p < 0.001. D, percentage of cells from experiments as in B with mCh-Rab13 on vesicles that exhibit perinuclear localization. Data are mean ± S.D., measuring a minimum of 20 cells/experiment from a minimum of three independent experiments. E, MCF10A cells expressing mCh-Rab13 T22N ΔC and internalized Alexa Fluor (AF) 647-labeled transferrin were imaged live. The arrows illustrate co-localization. The boxed region is magnified below over a time-lapse imaging series. Scale bars = 10 μm (top panel) and 1 μm (magnified panels). F, PC12 cells differentiated for 24 h with 50 ng/ml NGF and expressing mCh-Rab13 T22N-ΔC were imaged live. The boxed region is magnified. The arrows follow vesicle trafficking along a neurite. Scale bars = 10 μm (top panel) and 2.5 μm (magnified panel).