Figure 3.
Effect of Bam deletion mutants on sensitivity to NTZ. Strains BW25113ΔbamC/pMJ3 (PapCHis) (A), BW25113ΔbamB/pMJ3 (B), and BW25113ΔbamE/pMJ3 (C) were grown in the presence of the indicated concentrations of NTZ. WT bacteria containing vector only (pMON6235Δcat) or PapC (pMJ3) served as negative and positive controls for PapC expression, respectively. OM fractions isolated from normalized bacterial cultures were subjected to SDS-PAGE and Coomassie Blue staining to observe PapC and the overall OMP profile (top panels). Samples were also probed with anti-His tag antibody to visualize PapC (bottom panel). PapC levels were measured by densitometry of the anti-His tag blot, and percentage PapC levels were calculated relative to 0 μg/ml NTZ for each strain (D–F). Graphs show individual data points and means ± S.D. (error bars) from three independent experiments. ****, p < 0.0001 for comparison with 0 μg/ml NTZ.