FIGURE 3.

Immunoblotting of membrane preparations from wild type (WT), NuoI knock-out (KO), NuoI knock-out revertant (KO-rev), and site-specific NuoI mutants. E. coli membranes (15 μg of protein per lane) were loaded onto a 15% Laemmli SDS-polyacrylamide gel. After electrophoresis, the proteins were transferred onto nitrocellulose membranes, and Western blotting was carried out using the SuperSignal West Pico system. Antibodies specific to NuoB, NuoCD, NuoE, NuoF, NuoG, NuoI, NuoK, NuoL, and NuoM were used. Goat anti-rabbit IgG horseradish peroxidase conjugate was used as secondary antibody.