Fig 3. CoQ₁₀ production following addition of various concentration of Bz.

WT PR110 cells were pre-cultivated in 10 mL YES medium for 1 day. Cells at ~1×10⁵ cells/mL in YES media were cultivated for two days with rotation at 30°C in the presence of the indicated amount (μg/mL) of Bz, or without Bz. Gray bars show the CoQ₁₀ content per 50 mL of medium, and white bars show CoQ₁₀ normalized against cell number. Diamonds show cell number. Five micrograms of CoQ₆ was used as an internal standard. Data are represented as the mean ± SD of three measurements. (B) WT PR110 cells were pre-cultivated in 55 mL medium for 1 day. Yeast cells at an initial cell density of OD₆₀₀ 0.05 were cultivated in 1.5 L YES with 25 μg/mL of Bz, or without Bz, for 20 h with rotation at 30°C. Gray bars show the CoQ₁₀ content per 50 mL of medium, and white bars show CoQ₁₀ normalized against cell number. Diamonds show cell number. Five micrograms of CoQ₆ was used as an internal standard. Data are represented as the mean ± SD of two measurements. (C) From isolated mitochondria, lipids were extracted with hexane:methanol:isopropanol (5:2:1) and the amount of CoQ was measured by HPLC. Two micrograms of CoQ₆ was used as an internal standard for CoQ extraction. Protein concentration was measured by a Bio-Rad protein assay kit. (A−C) Asterisks on bars denote statistically significant differences (**p<0.01, *p<0.05) relative to samples from YES without Bz (Student’s t-test).