Figure 3

Accumulation of periplasmic LPS correlates with increased OMV production without alteration of Lpp crosslinking. (A) Relative fold OMV production in cultures of the indicated strains grown in LB overnight at 37°C was determined using FM4-64 and normalized to CFU as in Figure 1A. (B) Protein concentrations in periplasm preparations of the indicated strains grown ~16-18 hrs in LB at 37°C were determined by Bradford Assay. (C) Relative fold crosslinked Lpp in cultures of the indicated strains grown in LB to an OD₆₀₀ of ~ 0.4 at 37°C was determined as described in Figure 1B. (D) Relative fold of free Lpp in cultures of the indicated strains grown in LB at 37°C was determined as described in Figure 1C. (E) The lipid to protein ratio in the OMVs purified from cultures of the indicated strains grown in LB overnight at 37°C was determined by dividing the amount of lipid, measured using FM4-64, by the OMP concentration, measured by densitometry. (F) Relative fold OMV production in cultures of the indicated strains grown in LB overnight at 37°C was determined as described in Figure 1A. Error bars indicate SEM. *, p ≤ 0.05; n ≥ 3.