FIG 1.

Validation of PspA as a target for vaccination in biofilm dispersed bacteria. Biomass (A) and susceptibility (B) to gentamicin exposure (500 μg/ml for 3 h) of in vitro biofilms produced by wild-type and pspA-negative D39 bacteria following 48 h of growth over prefixed HREC. The bars show the geometric mean and 95% confidence interval of four biofilms. (C) Scanning electron microscopy images of WT and pspA-negative D39 biofilms formed on primary ciliated respiratory cells. (D) Colonization of D39 wild-type and pspA null pneumococci in mice at 48 h postinoculation. The dots represent individual mice, and the line represents the geometric mean of the data set. NP, nasopharyngeal. (E) Gene expression levels of pspA in biofilm and heat-dispersed EF3030 pneumococci. The graph presents the means and standard deviations. (F) Levels of PspA in whole-cell protein extracts from biofilm (BF) and released pneumococci from heat-dispersed (HD) and influenza (IAV)-treated biofilms grown over fixed or live HREC. (G) Quantification of pspA protein levels in biofilm (BF), heat-dispersed (HD), and influenza (IAV)-dispersed biofilm bacteria. Statistical significance of data in panels A, B, D, and E was determined using a Mann-Whitney test (**, P < 0.01; ns, not significant). Graphs in panels A, B, and E include data from at least two independent experiments. The graph in panel G represents the results from measurement of the bands shown in panel F.