Figure 7.

Redistribution of GBF1 from the 15°C peripheral compartments upon warm-up to 37°C is microtubule dependent. NRK cells were transferred to DMEM containing HEPES pH 7.4 and either 0.1% DMSO vehicle control or 5 μg/ml nocodazole (NOZ) and then incubated in a 15°C water bath for 2 h. Cells were either immediately fixed (0′) or quickly transferred to 37°C water bath and incubated for additional 1 min (1′), 3 min (3′), or 10 min (10′) before fixation. Coverslips were processed for IF by using polyclonal anti-GBF1 (e–h) or double-label IF (a–d and i–l) by using polyclonal anti- GBF1 (c and d, and I and j) and monoclonal anti-Man II (53FC3) (a and b, and k and l). Images obtained by standard epifluorescence microscopy are presented. Bar, 10 μm. Similar results were obtained in at least two independent experiments.