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. 2011 Sep;79(9):3683–3696. doi: 10.1128/IAI.01344-10

Fig. 1.

Fig. 1.

Subcellular localization of PdpE, IglG, and IglI in Francisella. C-terminal fusion proteins of PdpE and IglI (GSK tagged) and IglG (6×His tagged) were expressed from pKK289Km in the isogenic mutant background, resulting in ΔpdpE/pPdpE+(pMOL61) (A), ΔiglG/pIglG+(pMOL103) (B), and ΔiglI/pIglI+(pMOL59) (C) strains. Upon fractionation into soluble and membrane-associated fractions, Sarkosyl solubilization was used to further separate inner (Sarkosyl-soluble) and outer (Sarkosyl-insoluble) membranes. Protein fractions were separated by SDS-PAGE and analyzed using standard Western blot techniques and appropriate antisera. To detect PdpE and IglI, anti-GSK antiserum was used, while IglG was visualized using anti-His antibodies. Antibodies recognizing IglC, PdpB, or Tul4 were used as markers for soluble, inner membrane, and outer membrane fractions, respectively.