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Fig. 2 Pathophysiological indicators of infection in zebrafish larvae infected with the Mab ESX-3 mutant. (A) The percentage of larvae with abscesses after infection with R, R ΔeccC3, and R ΔeccC3::c at 3 days post-infection (dpi) and 5 dpi, and (B) representative images. Images were acquired in Widefield mode (Axiocam 712) with a TL LED lamp for brightfield and an LED-Module 590 nm for mScarlet and tdTomato. (C) The percentage of larvae with cords after infection with R, R ΔeccC3, and R ΔeccC3::c at 3 dpi and 5 dpi, and (D) representative images. Images were acquired in confocal mode with the 561 laser. Images are orthogonal projections of z-stacks. (E) the number of cords per larva and (F) the size distribution of cords at 3 dpi and 5 dpi. (A, C, E, and F) Data represent results from three pooled independent experiments, with an average of 24 larvae per condition per experiment. Relative il1b (G) and tnfa (H) gene expression measured by RT-qPCR at 1 h post-infection (hpi), 24 hpi, 72 hpi, and 120 hpi. PBS-injected embryos/larvae were used as controls. Data are from three pooled independent experiments, with 20 embryos/larvae per condition per experiment. (A, C, E, G, and H) Statistical analysis was performed per timepoint using ordinary one-way ANOVA with Tukey’s multiple comparisons test and after verifying the normality of the data.