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Figure 4


Fig. 4. Brefeldin A and ilimaquinone have differential effects on the vimentin cytoskeleton. SW13 v+ cells were treated in either the absence or presence of 10 µg/ml BFA or 40 µM IQ for either 30 minutes or 1 hour (vimentin 1 h panel only) at 37°C. Cells were then processed for immunofluorescence and stained with monoclonal antibodies directed against vimentin, GM130, the {gamma} subunit of AP-1 or the {delta} subunit of AP-3. Vimentin staining was visualized by epifluorescence microscopy, and GM130, AP-1 and AP-3 staining was visualized by confocal microscopy. BFA induced changes in vimentin architecture that became more pronounced at 1 hour. IQ did not induce similar changes, although cells were observed to round up, particularly with longer treatments. Both drugs resulted in fragmentation of the Golgi complex, as visualized by GM130 staining. Only BFA resulted in release of the adaptors AP-1 and AP-3 from the membrane. Bars, 20 µm. Insets are of cells indicated by an asterisk (*) and are magnified 3x.





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