Figure 4.

Automated Western blot analysis of the C‐terminally FLAG‐tagged version of BldO, expressed in trans from the native promoter in the bldO null mutant background.

Equal amounts (2.5 µg) of total protein were loaded for each sample and BldO‐FLAG was detected with anti‐FLAG antibody using the quantitative ‘Wes’ capillary electrophoresis and blotting system (ProteinSimple – San Jose, CA; see Supplementary Material). Wild‐type S. venezuelae expressing non‐FLAG‐tagged BldO was used as a negative control. Both the BldO FLAG‐tagged strain and wild type were grown in DNB medium. Top: DIC images of the culture at each time point. Middle: virtual Western blot. Bottom: quantitation of BldO levels (area under each peak; arbitrary units). All experimental samples were analysed in triplicate and the mean value and its Standard Error are shown for each sample.