FIG 3.

Both PDR1 and CDR1 are induced upon transcriptional inactivation of the camPr-ERG11 fusion. (A) The strain containing only the camR-VP16 transactivator (BVGC7) was grown to the mid-log phase and then treated with 50 μM camphor for the indicated lengths of time. Total RNA was prepared from each culture at each time point. Levels of the indicated mRNAs were determined by qRT-PCR. (B) Whole-cell protein extracts were prepared from the indicated BVGC7 cultures, and then the levels of Pdr1, Cdr1, Erg11-3× HA, and tubulin were determined by Western blotting with appropriate antisera. (C) Quantitation of Western blots prepared as described in the panel B legend. (D) BVGC9 (which contains both transactivator-responsive and camphor-responsive ERG11 alleles) was grown to the mid-log phase and treated with camphor to inactivate ERG11 expression for various times. RNA levels of ERG11, PDR1, and CDR1 were measured using qRT-PCR. (E) Whole-cell protein extracts were prepared from BVGC9 cultures treated with camphor for various times or left untreated. Western blot analyses were used to measure expression of the indicated proteins. (F) Quantitation of Western blots prepared as described for panel E.