FIG 7.

Loss of function of HAL9 results in upregulation of stress response protein HSP12. (A) Transcript levels of HAL9 and HSP12 were measured from untreated and treated (25 μM MMV688766) 16ABC parental cells by RT-qPCR and normalized to ACT1. Values were made relative to untreated 16ABC control sample. Error bars represent standard error of the mean (SEM) among technical triplicates. Significance was measured using a two-tailed unpaired t test with Welch’s correction: *, P < 0.05 compared to untreated. (B) Susceptibility of hsp12Δ, msn2Δ, msn4Δ, and msn2Δmsn4Δ mutants in the 16ABC and HAL9^A1543T mutant background. Each strain was assessed in 2-fold dose-response assay under shaking conditions at 30°C in YPD medium. Growth was measured after a 72-h incubation. (C) Susceptibility of pHSP12^OE16ABC mutant alongside empty-vector and parental (HAL9^A1543T) controls was tested in 2-fold dose-response assay under shaking conditions at 30°C in synthetic defined media (SD) to maintain plasmid selection. Growth was measured after a 72-h incubation. Growth for each strain was normalized to their respective drug-free controls and displayed using Java TreeView3; see color bar.