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. 2014 Dec;196(23):4012–4025. doi: 10.1128/JB.02034-14

FIG 2.

FIG 2

RpoE overexpression using a tetracycline-responsive expression system. (A) Effect of RpoE induction on the growth of 35000HP(pDG9) as assessed by OD measurements at 660 nm. An arrow indicates the point at which ATc was added to induce RpoE expression. (B) Effect of RpoE induction on the viability of 35000HP(pDG9) as assessed by quantitative culture. (C) RpoE induction as assessed by qRT-PCR analysis of rpoE transcripts following addition of ATc. (D) RpoE induction as assessed by Western blotting of RpoE expression following addition of ATc. Whole-cell lysates were prepared from 35000HP(pDG11) (lanes 1, 3, 5, and 7) and 35000HP(pDG10) (lanes 2, 4, 6, and 8) at 0, 5, 10, and 20 min after RpoE induction. RpoE expression was determined using a monoclonal antibody specific to the 3×-FLAG tag fused to the N-terminal end of RpoE immediately after the start codon. PAL, detected using the 3B9 antibody, served as a loading control. The data represent the means ± SD from four independent experiments. *, P < 0.05 and **P < 0.01.