Figure 10.

Nocodazole does not affect the spatial organization of SAC but perturbs the organization of the Golgi apparatus. In a and b, cells were labeled with C₆-NBD-GlcCer according to steps 1–4 of Fig. 1, to label SAC. In b, cells were subsequently incubated with 33 μM nocodazole at 4°C for 60 min. Note that nocodazole does not affect the appearance of the NBD lipid–labeled SAC. This was also the case when the nocodazole-treatment was done at elevated temperatures (van IJzendoorn, S.C.D., and D. Hoekstra, unpublished observations). In c and d, cells were labeled with 4 μM C₆-NBD-Cer at 4°C for 30 min, washed, and then incubated in back exchange medium at room temperature for 20 min. A typical Golgi apparatus labeling pattern was obtained (c). In parallel, such Golgi-labeled cells were subsequently treated with 33 μM nocodazole at 4°C for 60 min (d). Note the clearly different fluorescence pattern in the nocodazole-treated cells. Bar, 5 μm.