Fig. 4.
Analysis of membrane integration of SseF in dependence of its chaperone SscB (a) Salmonella ΔsscB, ΔsseF, ΔsseG triple mutants were grown under SPI-2-inducing conditions and complemented with SseFFLAG with or without the chaperone SscB from a rhamnose-inducible low-copy number plasmid. Crude membranes were prepared and treated with 8 M urea for 1 h at room temperature as indicated. SseF content was then analyzed by SDS PAGE, Western blotting and immunodetection with anti-FLAG and anti-YidC (inner membrane control) antibodies. b As in a but showing expression of indicated SseFFLAG mutants with increased hydrophobicity of the first TMS. A representative result of three independent experiments is shown. Abbreviation: CM: crude membranes