Fig. 2.
Comparative proteomics of L. pneumophila JR32, ∆lvbR and ∆lqsR biofilms and validation of differential P flaA expression.
A. Proteomics was performed with biofilms formed by ΔlvbR or ΔlqsR mutant strains and compared to the biofilms formed by the parental strain JR32. In the Venn diagrams all proteins are shown that are depleted or enriched in mutant biofilms (for details see Materials and Methods). Among the total of 1241 proteins identified, 342 were produced in concert depending on the presence of LvbR and LqsR (overlapping area; brackets: number of LvbR/LqsR‐dependent proteins, which accumulate in parallel (326, black) or reciprocally (16, grey).
B, C. P flaA ‐gfp expression in ∆lvbR and ∆lqsR mutant strains. Legionella pneumophila JR32, ∆lvbR or ∆lqsR strains harbouring the promoter‐reporter constructs P flaA ‐gfp (pCM009) or P flaA ‐gfp and P lvbR ‐lvbR (pRH022) were grown at (B) 25°C or (C) 37°C in AYE medium within microplates while orbitally shaking. GFP fluorescence (relative fluorescence units, RFU) at a gain of 50 and optical density at 600 nm (OD600) were measured over time using a microplate reader. The kinetics of the GFP fluorescence/OD600 values are shown. The data are means and standard deviations of a technical triplicate and representative of two (B) or three (C) independent measurements.