Fig 4.

Loss of Erg3 or Erg6a function leads to changes in azole and polyene susceptibility. (A, B, C) Amphotericin B (AMB, 8 mg/L), posaconazole (POS, 0.2 mg/mL), and isavuconazole (ISA, 8 mg/mL) susceptibility testing of wild type and ergosterol mutants on solid YPD media. Each species was assayed at its optimal temperature, 30°C and 26°C for M. circinelloides (Mci) and M. lusitanicus (Mlu), respectively. In (A), representative images of growth after 48 hours in all of the drug containing media and the control medium without drug (YPD). Two independently generated mutants were assayed for each gene deletion (Δ−1 and Δ−2). In (B, C), Mci (B) and Mlu (C) growth rates from each mutant across time (24 hour intervals) were determined as the percentage of growth area from the same strain cultured on YPD medium with and without drug. Individual values were determined either in six biological replicates for the wild type or in three biological replicates from two independently generated mutants (six total values, grouped together for simplicity as no significant differences were detected) and used to plot a smoothed curve and SD values as black lines. Strains were grouped by letters showing significant growth rate differences across the whole time-course (one-way ANOVA and Tukey HSD test, P-value ≤ 0.01).