Fig 4.

A. De-activation of complex I NADH dehydrogenase from parental and 29.9KD NADH dehydrogenase knockout strains in the presence or absence of itraconazole. SMP (50 mg/ml) from parental and the 29.9KD NADH dehydrogenase knockout were pre—incubated with 25 mM dNADH until the substrate was completely oxidised then the incubation was continued at 42°C for 10 minutes. The initial rates of the rotenone-sensitive dNADH:Q1 reductase activity were determined in the presence of 2 mM KCN, 2 mM MgCl₂, 100 mM dNADH and 50 mM Q1. The 100% activities were 0.315 and 0.12 mmol dNADH/minute per mg protein for the parental and 29.9KD NADH dehydrogenase knockout respectively. For addition of ITR 10μg itraconazole or a control amount of DMSO was added at time zero. B: Activity of complex I NADH dehydrogenase from parental and 29.9KD NADH dehydrogenase knockout strains during normal or hypoxic growth in liquid media. Activities were calculated as described above. Dissolved oxygen levels are shown as bars. O2+HYP: Oxygen level during growth of liquid cultures under mineral oil, O2normal; oxygen level in normal exposed liquid culture, 29.9; NADH dehydrogenase activity for the 29.9KD knockout in normal exposed liquid culture, Wild-type; NADH dehydrogenase activity for the parental strain in normal exposed liquid culture, 29.9+HYP; NADH dehydrogenase activity for the 29.9KD knockout during growth of liquid cultures under mineral oil, Wild-type; NADH dehydrogenase activity for the parental strain during growth of liquid cultures under mineral oil, Results represent the average of three biological experiments with three technical replicates each. Error bars represent standard deviation.