FIGURE 4.
Quantification of the different subunits of the F1F0-ATP synthase in RFY5-1 strain grown in the absence or presence of doxycycline. A and B, mitochondria isolated from RFY5-1 strain grown in YPGal medium in the absence or presence of doxycycline (Dox) (10 μg·ml-1) were prepared as described under “Experimental Procedures.” A, mitochondrial proteins (50 μg) were separated by Tris-Tricine SDS-PAGE, and the presence of the different subunits was revealed by Western blotting using appropriate antibodies. B, obtained signals were quantified. For each subunit, the relative quantity is the ratio between the signal obtained for mitochondria isolated from RFY5-1 strain grown in the presence of doxycycline (10 μg·ml-1) and the signal obtained for mitochondria isolated from the same strain grown in the absence of the antibiotic and normalized to the ratio obtained for porin. This ratio is expressed in percentage (relative quantity (%) = 100 × ((signal RFY5-1 + Dox for subunit x)/(signal RFY5-1-Dox for subunit x)) × ((signal RFY5-1 + Dox for porin)/(signal RFY5-1-Dox for porin))). Measurements were obtained from four different mitochondrial preparations.