Figure 2.

Depletion of HSP90 increases sensitivity to fluconazole. (A) Wild-type and CTR4-2p-HSP90 strains were 10-fold serially diluted and spotted on YPD agar and YPD agar supplemented with 25 µM CuSO₄ with or without 0.5 µg/ml amphotericin B or 2 µg/ml fluconazole. Agar plates were incubated at 37°C and imaged after 2 days. (B) Wild-type and CTR4-2p-HSP90 strains were spread on YPD agar with or without 5 µM CuSO₄, and an ETEST strip of amphotericin B or fluconazole was placed in the center of the plate to test antifungal drug susceptibility. Plates were imaged after incubation at 37°C for 6 days. (C) Wild-type cells were inoculated at 1 × 10⁶ CFU in liquid YPD medium in the absence or presence of fluconazole, radicicol, HSP990, or a combination as indicated. Cells were cultured at 37°C and plated on YPD agar medium at 0, 4, 8, and 24 h postinoculation for CFU determination. Error bars represent standard deviation of the mean for technical triplicates. (D) Wild-type and CTR4-2p-HSP90 strains were inoculated at 1 × 10⁶ CFU in liquid YPD medium in the absence or presence of fluconazole, CuSO₄, or a combination as indicated. Cells were cultured at 37°C and plated on YPD agar medium at 0, 4, 8, and 24 h postinoculation for CFU determination. Error bars represent standard deviation of the mean for technical triplicates.