FIG. 6.

(A) Northern blot analysis of UPC2 strains. BWP17 and UPC2 deletions strains were grown in minimal medium in the absence or presence of 5 μg of fluconazole/ml for 6 h to an OD of 0.8 to 1.0. Total RNA was harvested, run on an agarose gel, blotted onto a nitrocellulose membrane, and then probed with radiolabeled oligonucleotides specific for each gene (see Materials and Methods). A probe of the ACT1 housekeeping gene served as the loading control for each blot; all strains had equivalent levels of ACT1 mRNA (data not shown). (B) Quantification of Northern blot results. mRNA levels for BWP17 and UPC2 deletion strains were measured by using a Storm Phosphorimager. After normalizing each band to its ACT1 loading control, a comparison of the relative intensities between strains grown with and without drug showed a 6- to 16-fold increase in the expression of ERG2 (light bars) and ERG11 (dark bars) in the presence of 5 μg of fluconazole/ml. UPC2 expression changes could not be measured, as signal in the no-drug lanes was not significantly above background.