FIG. 4.

LolCDE overexpression suppresses the growth inhibition by LolA(I93C/F140C). (A) An overnight culture of C43(DE3) cells harboring both pSWIF (I93C/F140C) and pCOLA-CDE (LolCDE) was diluted 200-fold with fresh medium containing 0 (closed circles), 0.05 (closed triangles), 0.25 (closed diamonds), or 1 (closed squares) mM IPTG, followed by cultivation for 2 h. Arabinose (0.2%) was then added to induce I93C/F140C. As controls, growth in the absence of arabinose with (open squares) or without (open circles) 1 mM IPTG was also examined. (B) Total cell lysates were prepared from equivalent amounts of cells at the end of the experiments shown in panel A and then subjected to SDS-PAGE and Western blotting with anti-FLAG peptide and anti-LolC and anti-DegP antisera. The band indicated by an asterisk below the His-LolC band represents chromosomally encoded wild-type LolC, and those indicated by asterisks below DegP represent those mentioned in the legend to Fig. 2E. As a control, C43(DE3) cells harboring pSW77 (wild-type LolA) and pCOLA-CDE (LolCDE) were also examined as indicated.