Figure 4.

Effect of mβCD and mβCD/chol on the Rab9-independent pathways to the Golgi apparatus. (A) Representative example of the resulting bands seen after fluorography in a given experiment. The HeLa-TetOn/Rab9S21N cell line was grown with or without doxycycline (2μg/ml) for 18 h in order to express mRab9. The cells were then washed with sulfate-free DME medium, and incubated for 4 h in the presence of Na₂³⁵SO₄ before addition of 5 mM mβCD or mβCD/chol. After 30 min, ricin sulf-2 was added, and the incubation continued for 2 h. The sulfated ricin molecule was then harvested and analyzed as described in Materials and Methods. The intensities of the resulting bands were determined by densiometric quantitation using ImageQuant 5.0. (B) Summary from two independent experiments.