Glutamate dehydrogenase (Gdh2)-dependent alkalization is dispensable for escape from macrophages and virulence of Candida albicans
Fig 2
C. albicans Gdh2 localizes to the mitochondria and environmental alkalization requires mitochondrial function.
(A) Gdh2-GFP co-localizes with the mitochondrial marker MitoTracker Deep Red FM (MTR). YPD grown cells expressing GDH2-GFP (CFG273) were harvested, washed, grown in SED (0.2% glucose) or YNB + CAA at 37°C for 24 h, and stained with 200 nM MTR prior to imaging by differential interference contrast (DIC) and confocal fluorescence microscopy; the scale bar = 5 μm. (B) Wildtype cells (SC5314) from overnight YPD cultures were washed and then diluted to either OD600 ≈ 0.1 (top panel) or ≈ 5 (bottom panel) in liquid YNB+CAA with the indicated concentrations of mitochondrial complex III inhibitor antimycin A. Cultures were grown at 37°C under constant aeration for 24 h and 2.5 h, respectively, and photographed. To assess viability after Antimycin A treatment, inhibited cells from 24 h old culture (top panel) were harvested, washed, and then resuspended in fresh YNB+CAA media and incubated for 24 h (up to 48 h) at 37°C (middle panel). Images are representative of at least 3 independent experiments.