FIGURE 6.

Sumoylated wild type and GTP-bound, but not GDP-bound rab17, are membrane associated. A, live WIF-B cells expressing myc-tagged wild type rab17 were extracted with 0.1% Triton X-100 in Hepes buffer at 37 °C for the indicated times. The buffer with extracted cellular contents was collected and immunoblotted for rab17, α-tubulin, or HDAC6 as indicated. Arrows mark the two rab17 species. B, WIF-B cells expressing wild type myc-tagged rab17 were fractionated as described in the legend to Fig. 5 and fractions were immunoblotted for rab17, APN, and HDAC6 as indicated. Numbers on the right indicate the percent total of either species detected in the final membrane pellet. C, WIF-B cells expressing FLAG-tagged Q77L or N132I rab17 were detergent pre-permeabilized, fixed, and stained with anti-FLAG antibodies. Arrows mark the subapically located Q77L rab17. Asterisks mark bile canaliculi in panels c and d. Bar = 10 μm.