FIG 4.

Effects of dead YPC1 alleles, australifungin, and myriocin on cells having reduced IPC synthase activity. (A) Serial dilutions of cells overexpressing either WT or two functionally dead forms of YPC1 (denoted * and **) were plated on plates containing AbA or Doxy. (B) tetAUR1 containing hemagglutinin (HA)-tagged YPC1, empty vector, or the two dead alleles of YPC1 behind the GAL1 promoter were grown in galactose for 20 h, and extracts corresponding to 1 OD₆₀₀ unit of cells were Western blotted using anti-HA. (C) Upper two rows, serial dilutions of tetAUR1 were spotted on YPD and YPD supplemented with 0.25 μg/ml Doxy, 0.03 μg/ml AbA, 1.50 μg/ml australifungin, or combinations thereof at the same concentrations. Lower two rows, cells were also spotted on media supplemented with 0.50 μg/ml Doxy, 0.01 μg/ml AbA, or 0.05 μg/ml myriocin or combinations thereof at same concentrations. Plates were incubated for 2 days at 30°C. (D) Tenfold dilutions of vma mutants in the BY4742 (WT) or tetAUR1 background were spotted onto YPD plates with the indicated concentrations of AbA and NAC. (E) To demonstrate the efficacy of NAC in antagonizing the deleterious effects of ROS, the ROS-hypersensitive sod1Δ cells, lacking cytosolic superoxide dismutase, were included to show that the growth inhibition caused by paraquat, an oxidant, can be antagonized by NAC.