Journal: microLife
Article Title: The opposite effects of stringent response on phage infection of Pseudomonas putida
doi: 10.1093/femsml/uqaf048
Figure Lengend Snippet: (p)ppGpp affects P. putida cell morphology and permeability in liquid LB medium. Pseudomonas putida PaW85 and its relA deletion (ΔrelA) and (p)ppGpp 0 derivative (ΔrelAΔspoT) populations were compared in late exponential phase (OD1), stationary phase (24 h), and (in c and d) late stationary phase (48 h). (a) Microscopy images of LB-grown cultures. White arrows indicate elongated cells and black arrows the aggregates present in stationary phase (p)ppGpp-deficient cultures. (b) Box plots with Tukey whiskers and individual outliers of single cell sizes (area) analysed from microscopy images using MicrobeJ. Brown-Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test with individual variance. (c) Population differences from flow cytometry analysis: cells stained with propidium iodide (PI), Syto 9, and cells with reduced fluorescence intensity are brought out. Presented are means with SD ( N = 3). Mutants were compared to wild-type in each time point with ordinary two-way ANOVA with Tukey’s multiple comparisons test with single pooled variance. (d) Flow cytometry analysis of cells stained with membrane potential indicator dye DiOC 2 (3). Geometric means of red and green fluorescence of the single cell population were analyzed. Higher red/green ratio indicates higher membrane potential. Means ± SD ( N = 3). Ordinary two-way ANOVA with uncorrected Fisher’s Least Significant Difference (LSD) test with a single pooled variance. * P -value <.05; ** P -value <.005; *** P -value < .001; **** P -value <.0001; RFU—relative fluorescence unit.
Article Snippet: Fluorescent dyes DiOC 2 (3) (3,3'-Diethyloxacarbocyanine Iodide; final concentration 30 μM) and DiSC 3 (5) (3,3'-Dipropylthiadicarbocyanine Iodide; final concentration 5 μM) (Thermo Fisher Scientific) were used to estimate membrane potential.
Techniques: Permeability, Microscopy, Flow Cytometry, Staining, Fluorescence, Membrane
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