Strains harboring a 4xCDRE-lacZ construct were cultured in the absence or presence of 64 μg/ml fluconazole, and with or without 10 μM geldanamycin, 1 μg/ml FK506 (in DMSO, A. G. Scientific), or 20 μg/ml beauvericin. Strains were grown in synthetic defined (SD) media at 25°C overnight and diluted the next day to OD600 0.01 ± inhibitors geldanamycin, FK506, or beauvericin. Cultures ± inhibitors were again incubated overnight in SD at 25°C and diluted the next day to OD600 0.3 ± inhibitors in the presence or absence of fluconazole for five hours. Strains harboring the HSP70-lacZ construct were treated with 10 μM geldanamycin or 10 μg/ml beauvericin. Strains were grown in YEPD at 30°C overnight and diluted the next day to OD600 0.01 ± inhibitors geldanamycin or beauvericin. Cultures ± inhibitors were again incubated overnight in YEPD at 30°C and diluted the next day to OD600 0.2 ± inhibitors for four hours. Post-treatment sample processing and analysis was conducted as previously described10 (link).
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Measuring Stress Response Pathways
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Corresponding Organization :
Other organizations : University of Toronto, University of Arizona, Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Duke University, Duke University Hospital, Duke Medical Center, Yonsei University, University of Milano-Bicocca, Heinrich Heine University Düsseldorf, University of Geneva, University of Wisconsin–Madison, University of British Columbia
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Variable analysis
independent variables
- Presence or absence of 64 μg/ml fluconazole
- Presence or absence of 10 μM geldanamycin
- Presence or absence of 1 μg/ml FK506
- Presence or absence of 20 μg/ml beauvericin
- Presence or absence of 10 μM geldanamycin (for HSP70-lacZ construct)
- Presence or absence of 10 μg/ml beauvericin (for HSP70-lacZ construct)
- Expression of the 4xCDRE-lacZ construct
- Expression of the HSP70-lacZ construct
- Growth in synthetic defined (SD) media at 25°C
- Growth in YEPD media at 30°C
- Not specified
- Strains cultured in the absence of any inhibitors
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