Fig. 6: Overexpression of insA restores triazole susceptibility to hmg1 overexpression mutants but does not resolve resistance due to SSD mutation.
From: A secondary mechanism of action for triazole antifungals in Aspergillus fumigatus mediated by hmg1
A Schematic of promoter replacement mutations for insA in the hgm1pHspA and hmg1S305P genetic backgrounds. Illustration prepared using BioRender.com. B RT-qPCR analyses were performed in each background to ensure that the genetic manipulation at the hmg1 locus of the parental strain did not alter insA expression and to confirm that the promoter replacement resulted in overexpression of insA. Averaged data represent the mean ± standard deviation. n = three biologically independent samples and statistical analyses were performed by One-Way Anova followed by an unpaired T test with significance set at 0.05 and degrees of freedom equal to 10 for individual comparisons.Radial growth assays of the parental control, hmg1 overexpression (hmg1pHspA), hmg1/insA double overexpression (hmg1pHspA/insApHspA), hmg1 SSD (hmg1S305P), and hmg1 SSD/insA overexpression (hmg1S305P/insApHspA) mutants to assess susceptibility changes are shown for rosuvastatin (C), terbinafine (D), and voriconazole (E). Conidia (5 × 103) from each strain were spot inoculated onto RPMI media (0.2% glucose, pH 7.0) containing the indicated amount of drug and plates were cultured for 72 hours at 37 °C. Source data are provided as a Source Data file.
