Figure 2.

Effect of the identified drugs on WB-12/aac2^M114P phenotypes. Wild type WB-12/AAC2, null mutant (aac2Δ), and WB-12/aac2^M114P mutant strains with or without the supplementation of active compounds were grown in YP medium supplemented with 0.6% glucose at 28 °C. (A) Oxygen consumption rate analysis. The highest and the lowest effective concentrations are reported for each molecule. Values were normalized to the values of the wild type strain and represented as the mean of at least four independent experiments ± SD. (B) Oxygen consumption rate under normal (yellow bars) and uncoupled state condition using the mitochondrial uncoupler CCCP (pink bars). Values were normalized to values of the wild type strain without CCCP and represented as the mean of three independent experiments ± SD (C) Mitochondrial membrane potential (MMP) was measured by the uptake of the fluorescent dye DiOC₆. Values were normalized to the values of the wild type strain and represented as the mean of three independent experiments ± SD. (D) ROS production was determined as the percentage of fluorescent cells using DHR123 probe. In addition to the shown concentrations, halved concentrations were also tested, and similar results were obtained. The antioxidant agent N-acetyl-cysteine was used at two different concentrations (15 mM and 30 mM) as a positive control. All values are means of three independent experiments ± SD. Statistical analysis was performed using an unpaired, two-tailed Student’s t-test, comparing treated (blue bars) versus untreated (red bar) for (A,C,D), and comparing w/o CCCP (yellow bar) versus with CCCP (pink bar): * p < 0.05; ** p < 0.01; *** p < 0.001. The same amount of DMSO (compound vehicle) was added to the untreated mutant (red bar). OTI = otilonium bromide, PER = pergolide mesylate, TRI = trifluoperazine 2HCl, SER = sertraline HCl, BEN = benzydamine HCl.