Zebrafish survival to P. aeruginosa infections. Kaplan–Meier survival curves of zebrafish embryos microinjected in the pericardial cavity as described in Section 4.2, with increasing doses of PA14 wild-type or PBS alone (ctrl). The survival was calculated as a percentage of the total injected embryos (n = 15) for each treatment group and was monitored at post infections time intervals (hpi). Data were obtained from one experiment representative of three independent experiments.

Competition assays of P. aeruginosa strains in zebrafish embryos. (A) Competition between zrmA (strain A) and wild-type (strain B). (B) Competition between znuA (strain A) and wild-type (strain B). (C) Competition between znuAzrmA (strain A) and wild-type (strain B). (D) Competition between znuAzrmA (strain A) and znuA (strain B). Each data point represents the competitive index calculated, as described in Section 4.3, on a homogenate of 6 pooled animals. A horizontal straight line indicates the median CI value calculated on the CI values of independent experiments. Statistical analyses were performed by the Student’s t-test (ns, not significant).

Expression levels of slc30 genes from zebrafish infected with P. aeruginosa. RT-PCR was performed on mRNA extracted from infected zebrafish at 6, 24, and 48 hpi. The dotted line at Y = 1 is the expression level of the genes in uninfected animals (control). Data are means ± SD of three independent experiments, and statistical analyses were carried out by two-way ANOVA and Tukey’s multiple comparison test. Asterisks show statistical differences versus uninfected zebrafish (**** p < 0.0001; *** p < 0.0005; * p < 0.05), and hash signs show statistical differences between 24 hpi and 48 hpi for slc30a4, and between 6 hpi and 48 hpi for slc30a7 (# p < 0.05).

Expression levels of P. aeruginosa genes from infected zebrafish. RT-PCR was performed on bacterial mRNA extracted from zebrafish at 6, 24, and 48 hpi. The dotted line at Y = 1 represents the expression level of the genes in bacteria grown in LB medium (control). Data are means ± SD of three independent experiments, and statistical analyses were performed by ordinary one-way ANOVA and Bonferroni’s multiple comparison test. Asterisks show statistical differences versus bacteria grown in LB medium (**** p < 0.0001; *** p < 0.0005; * p < 0.05), hash signs show statistical differences between samples at the different hpi (#### p < 0.0001; ## p < 0.001; # p < 0.05); nd: not detected.