Fig. 1: Increased cyclic-di-GMP levels downregulate T3SS assembly and activity, and upregulate H1-T6SS assembly and activity. | Communications Biology

Fig. 1: Increased cyclic-di-GMP levels downregulate T3SS assembly and activity, and upregulate H1-T6SS assembly and activity.

From: Coordination of virulence factors and lifestyle transition in Pseudomonas aeruginosa through single-cell analysis

Fig. 1

A Fraction of T3SS-positive bacteria in different c-di-GMP backgrounds. Data is shown as a fraction of cells with EGFP-SctQ foci; exact numbers are shown above each bar. Samples were taken from >10 different fields of view in 3 different experiments. Statistical analysis was done via χ2 test, ns, non-significant (the precise numbers of all p-values are provided in Suppl. Data 1). B Number of T3SS per bacterium in different c-di-GMP backgrounds. Data is shown as the number of EGFP-SctQ foci per T3SS-positive cell, with each data point representing the calculation from one field of view. Samples were taken from >10 different fields within 3 different experiments. Statistical analysis was done via ANOVA with multiple comparisons to WT, ****, p < 0.0001; ns, non-significant. C T3SS effector secretion in different c-di-GMP backgrounds examined by label-free quantitative mass spectrometry. ExoS/Y/T are the three T3SS effectors in PAO1; protein intensities (reflecting the abundance of a specific protein in a sample) in the supernatant are compared in different c-di-GMP backgrounds. Data is shown as log2 mean intensity of effector proteins for the different background strains. Bars indicate the mean value. Biological replicates, n = 3. Statistical analysis was done via ANOVA with multiple comparisons to WT, ***, p < 0.001; ns, non-significant. D Fraction of H1-T6SS-positive bacteria in different c-di-GMP backgrounds. Data is shown as a fraction of cells with H1-T6SS assembly (TssB1-mCherry foci); exact numbers are shown above each bar. Samples were taken from >10 different fields of view in 3 different experiments. Statistical analysis was done via χ2 test, **, p < 0.01; ****, p < 0.0001. E H1-T6SS killing efficiency test in different c-di-GMP backgrounds. Indicated strains were used as predators, while a YFP-labeled H1-T6SS effector-immunity pair deletion strain (Δtse6tsi6) was used as prey. Data is shown as background-corrected prey fluorescence. Data points are shown as an endpoint value of a 24-h killing assay. Biological replicates, n = 3. Statistical analysis was done via ANOVA with multiple comparisons to WT, **, p < 0.01; ****, p < 0.0001.

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