Fig. 3

Shigella-trained macrophages produce more ROS (A) Schematic of the zebrafish immune training model in the HBV and re-infection in the duct of Cuvier. (B and C) Survival rates (B; N = 3 with >14 larvae per experiment) and bacterial burden (C) of naive and Shigella-trained (1.5 × 103 ± 2.5 × 102 CFUs) larvae upon systemic secondary infection (PBS 2.4 × 104 ± 8.2 × 103 CFUs and Shigella 1.7 × 104 ± 9.9 × 103 CFUs). (D) Quantification of tnfa:GFP expression in naive and Shigella-trained macrophages at 3 hp2i (N = 3 with >20 larvae per experiment). Single dots indicate Shigella-infected macrophages. (E) Quantification of LIVE/DEAD staining internalized by macrophages in naive and Shigella-trained larvae at 3 hp2i (N = 4 with >20 larvae per experiment). (F) Quantification of CellROX in naive and Shigella-trained macrophages at 3 hp2i (N = 3 with >20 larvae per experiment). Single dots indicate Shigella-infected macrophages. ns, non-significant; ∗∗∗∗p < 0.0001. Error bars represent standard deviation (SD). (B) Log rank (Mantel-Cox) test. (C) Two-way ANOVA with Sidak’s multiple comparisons test. (D–F) One-way ANOVA with Tukey’s multiple comparisons test). Percentages indicate population with normalized values ≥ 1.