Fig. 4

Localisation of Rpf-mCherry/EGFP fusions by western blot: culture supernatants. SDS-PAGE gels were loaded with 20 μl of concentrated culture supernatants (previously precipitated and resuspended in 40 μl of loading buffer; see Methods). The letters A-E correspond to samples from RpfA-E-mCherry/EGFP-producing cultures, respectively. mCh and EGFP refer to the control strains expressing fluorophore alone. The molecular weight ladder (L) values in kDa are shown on the left of each gel. a The western blots performed with culture supernatants of RpfA-mCherry and RpfE-mCherry producing strains present bands at the expected heights for the fusion proteins, as seen in soluble and precipitated extracts (Fig. 3). b In the western blots performed with culture supernatants of Rpf-EGFP producing strains, none of the samples present bands at the expected heights, indicating that probably none of the fusion proteins is being secreted to the extracellular medium. Hypothetical molecular weight of the fusion proteins: see Fig. 3 legend