Figure 4

trib1 overexpression is host protective against Mm infection. (A) Stereo-fluorescence micrographs of Mm mCherry infected 4 dpi larvae after injection at the single-cell stage with dominant active hif-1α (DA1), dominant negative hif-1α (DN1), trib1, trib2, trib3 RNAs and phenol red (PR) as a negative vehicle control. DA1 and DN1 are RNA controls with DA1 having previously been shown to reduce infection levels (Elks et al., 2013). Scale bar = 200 µm. (B) Bacterial burden of larvae shown in (A). Data shown are mean ± standard error of the mean (SEM), n = 76–77 in trib1, n = 86–89 in trib2, and n = 43–95 in trib3, accumulated from three independent experiments for each trib gene. Statistical significance determined via one-way analysis of variance (ANOVA) with Bonferroni’s multiple comparisons. p values shown are: **p < 0.01 and ***p < 0.001. (C) Stereo-fluorescence micrographs of Mm mCherry infected 4 dpi larvae after injection with tyrosinase (control), trib1 and trib3 CRISPR guides (CRISPants). Scale bar = 200 µm. (D) Bacterial burden of larvae shown in (C). Data shown are mean ± SEM, n = 87–90 fish accumulated from three independent experiments. Statistical significance determined via one-way ANOVA with Bonferroni’s multiple comparisons. p values shown are: ***p < 0.001. (E) Bacterial burden of trib1−/− stable knockout larvae compared to wildtype (trib1+/+) siblings. Data shown are mean ± SEM, n = 82–114 fish accumulated from four independent experiments. Statistical significance determined via an unpaired t-test. p values shown are: *p < 0.05.