<i>In vitro</i> phenotypes of glyoxalase mutants.

(A) Plaque area measured 3 days post-infection as a percentage of wild-type. Mean and standard error of the mean (SEM) pooled from three independent experiments is shown (n = 30). (B) Broth growth curve of indicated L. monocytogenes strains grown in BHI medium at 37°C with shaking. Mean and standard deviation of three independent experiments is shown. BMM’s were infected at an MOI of 0.25 with L. monocytogenes without treatment (C) or treated with PAM3CSK4 (D) and intracellular CFU were enumerated at different time points. Data are mean and SEM of three technical replicates of three independent experiments. For both panels, statistical significance is shown for ΔgloA compared to wild-type L. monocytogenes. (E) Sensitivity to MG (20% v/v) as measured by growth inhibition in a disk diffusion assay as percentage of wild-type. Data are mean and SEM of at least three independent experiments. For all experiments p values were calculated comparing to the wild-type bacteria using an unpaired Student’s t- test; *P < 0.05, **P < 0.01, ***P < 0.001, **** indicates P <0.0001.