Fig 8. Western blotting of Coq4, Coq8, and Dlp1 under various Bz concentrations.
(A) Coq4, Coq8, Dlp1, and Cdc2 as a loading control for whole cells were analyzed by western blotting. Target proteins are indicated on the right. The concentrations of Bz in each lane are shown at the top. For the pre-culture, PR110 yeast cells were cultivated in 10 mL YES for 1 day. Yeast cells were cultivated in 55 mL YES at an initial cell density of 1×105 cells/mL and cultivated for 48 h with rotation at 30°C. (B) Mitochondria were isolated as described in Materials and Methods. A 5 μg protein sample from the whole cell extract (WCE) or purified mitochondria (Mito)-enriched samples were used (right panel). (C) WT (PR110), Δppt1 (KH2), and Δcoq4 (KH4) strain with 100 μg/mL of Bz were cultivated in the YES media containing ~1×105 cells/mL for two days with rotation at 30°C. Then, Coq4 and Cdc2 proteins were detected by western blotting as described above. The amount of proteins was quantified by Image J.