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. 2018 Feb 5;164(3):338–348. doi: 10.1099/mic.0.000610

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Fig. 4. — Expression of the YscE-6xHis (G35K G36H D37K) mutant in ΔyscE Y. pestis stabilizes YscF as part of a cytoplasmic YscEFG complex, but YscF is not secreted. (a) A Y. pestis ΔyscE mutant complemented with pYscE-6xHis or pYscE-6xHis (G35K G36H D37K) was grown in the presence (+) or absence (−) of 2.5 mM CaCl₂, and the cell pellets and culture supernatants were analysed by SDS-PAGE and immunoblot analysis with antibodies specific for YopN, YscF and penta-His. The YscF stabilized by YscE-6xHis (G35K G36H D37K) is marked by an asterisk. (b) The presence of a cytoplasmic YscEFG complex was determined by BS3 crosslinking, SDS-PAGE and immunoblot analysis with antibodies specific for YscF. The location of the YscEFG complex in the parent (WT) and yscE deletion strains expressing YscE-6xHis or YscE-6xHis #11 is shown by an arrowhead.