Fig. 3: QS dependent increase in levels of elastase and rhamnolipids drives hypervirulence of PAO1 ∆mexEFoprN. | Nature Communications

Fig. 3: QS dependent increase in levels of elastase and rhamnolipids drives hypervirulence of PAO1 ∆mexEFoprN.

From: Evolutionary loss of an antibiotic efflux pump increases Pseudomonas aeruginosa quorum sensing mediated virulence in vivo

Fig. 3

a Fold change in lasB gene expression in PAO1 and PAO1 ∆mexEFoprN measured by RT-qPCR. Data show mean ± SEM, n = 3 independent experiments. Statistical significance analyzed by two-sided unpaired t-test. b Fold change in rhlA gene expression in PAO1 and PAO1 ∆mexEFoprN measured by RT-qPCR. Data show mean ± SEM, n = 3 independent experiments. Statistical significance analyzed by two-sided unpaired t-test. c Elastase activity determined from the supernatants of PAO1 and PAO1 ∆mexEFoprN using a fluorogenic substrate. Fluorescence has been normalized to total protein levels in the culture supernatants. Data show mean ± SEM, n = 6 independent experiments. Statistical significance analyzed by two-sided unpaired t-test. d Rhamnolipid production in PAO1 and PAO1 ∆mexEFoprN using cetyltrimethylammonium bromide (CTAB) methylene blue agar plates. A halo (white arrow) indicates rhamnolipid production. e Bacterial CFU enumerated from the whole lung lysate of PAO1 (n = 21), PAO1 ∆mexEFoprN (n = 16), PAO1 ∆mexEFoprN∆lasB (n = 5), PAO1 ∆mexEFoprN∆rhlA (n = 7) or PAO1 ∆mexEFoprN∆lasB∆rhlA (n = 6) infected C57BL/6 mice at 24 hpi. Data show mean ± SEM. Statistical significance analyzed by ANOVA Kruskal-Wallis test. f Percentage of PAO1 (n = 21), PAO1 ∆mexEFoprN (n = 16), PAO1 ∆mexEFoprN∆lasB (n = 5), PAO1 ∆mexEFoprN∆rhlA (n = 7) or PAO1 ∆mexEFoprN∆lasB∆rhlA (n = 6) infected C57BL/6 mice with viable bacteria in the liver at 24 hpi. Statistical significance analyzed by Fisher’s exact test. n indicates the number of mice/group. g Total PQS levels quantified in PAO1 and PAO1 ∆mexEFoprN by thin-layer chromatography (TLC). Data show mean ± SEM, n = 7 independent experiments. Statistical significance analyzed by two-sided unpaired t-test. h Intracellular PQS levels quantified in PAO1 and PAO1 ∆mexEFoprN quantified by TLC. Data show mean ± SEM, n = 7 independent experiments. Statistical significance analyzed by two-sided unpaired t-test. i Fold change in lasB gene expression in PAO1 ∆mexEFoprN and PAO1 ∆mexEFoprN∆pqsA measured by RT-qPCR. Data show mean ± SEM, n = 3 independent experiments. Statistical significance analyzed by two-sided unpaired t-test. j Fold change in rhlA gene expression in PAO1 ∆mexEFoprN and PAO1 ∆mexEFoprN∆pqsA measured by RT-qPCR. Data show mean ± SEM, n = 3 independent experiments. Statistical significance analyzed by two-sided unpaired t-test.

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