Figure 7.

Calcineurin is required for azole and echinocandin tolerance in C. glabrata. (A, B) Calcineurin mutants are hypersensitive to azoles and echinocandins. Cells were grown overnight in YPD at 24°, 5-fold serially diluted, spotted onto YPD medium containing fluconazole (FLC), posaconazole (PSC), ketoconazole (KTC), micafungin (MCF), caspofungin (CSF), or anidulafungin (ANF), and incubated at 24° for 48 hr. (C, D) Disk diffusion assays were used to determine fluconazole and micafungin susceptibility of wild-type and mutant strains. Cells were grown overnight at 24°, and 0.1 OD₆₀₀ (in 100 μl) was spread on the surface of YPD medium. A disk containing 20 μg of fluconazole (C) or 50 ng of micafungin (D) was placed on the surface of the medium. The plates were incubated at 24° for 24 hr. (E, F) Time-killing curve of wild-type and cna1 (YC98), cnb1 (YC193), and crz1 (YC182) mutants in YPD medium ± fluconazole (E) or micafungin (F). The experiments were performed at 24°. The data are represented as means ± SD from triplicate experiments.