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Fig. 8. Filipin specifically stained yeast sterols. (A) Wild-type ERG9 (CBY858) and erg9{Delta} PMET3-ERG9 (CBY745) cells were treated with filipin. The left panels show cell morphology by DIC, and the right panels show the corresponding filipin staining observed by fluorescence microscopy. After addition of methionine, filipin fluorescence in erg9{Delta} PMET3-ERG9 cells was considerably less intense than in wild-type ERG9 cells. Photographs were taken with equivalent exposures, although with longer exposures background filipin fluorescence was detectable in the ERG9-repressed cells. (B) Sterol distribution in both arv1{Delta} (CBY994) and wild-type ARV1 (CBY858) cells were also examined using filipin fluorescence. In 41% of arv1{Delta} cells (52 out of 136 cells), fluorescence was observed within the cells whereas in wild-type cells, only 10% of cells (10/96) exhibited a comparable staining pattern. Arrows indicate filipin fluorescence associated with internal membranes.





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