Fig. 6

Loss of PFD2 results in mild deficiencies in respiratory chain complexes. A–E Yeast cells were grown in complete synthetic medium that contained glycerol at 25 °C and shifted to 53 °C for 1 h where indicated. Mitochondria were isolated for both growth conditions. A–C Mitochondrial extracts were separated on native gel and analyzed by Western blot using specific antibodies. Experiments were performed in three biological repetitions. D, E Mitochondrial extracts were separated on SDS-PAGE and analyzed by Western blot using specific antibodies. Blots shown in D serve as loading control for native gel shown in A. Blots shown in E serve as loading control for native gels shown in B and C. F Quantification of mitochondrial protein levels. The data are expressed as the mean ± SEM. n = 3. G–I Yeast cells were grown in galactose-containing medium at 25 °C and shifted to higher temperature as indicated. Total protein extracts were separated on SDS-PAGE and analyzed by Western blot using specific antibodies. G Yeast cells were treated with 40 µM CCCP or an equivalent volume of DMSO (solvent) for 15 min. Cells were harvested before treatment (-) or after treatment (DMSO or CCCP). WT, wild type. Uncropped blots are presented as source data in the Additional file 13