Fig. 3

In vivo efficacy of TriSLas against M. marinum-infected embryonic zebrafish. (a) Schematic of the in vivo efficacy protocol showing the timeline of zebrafish embryo infection (200–250 colony-forming units (cfu) M. marinum expressing Wasabi), the exposure period to different concentrations of TriSLas (daily changes), and compound washout. hpf: hours post-fertilization; dpi: days post-infection; FPC: Fluorescence pixel count. (b–d) Survival curve ofM. marinum-infected zebrafish embryos monitored over the 10-day period post-infection, with and without 4-day treatment of (b) compound 11, (c) compound 12, and (d) compound 13. Survival curves are the cumulative results of three experiments covering more than 25 infected zebrafish per group. Statistics to compare treatment groups were performed using the log rank (Mantel–Cox) statistical test (*P < 0.05, **P < 0.01, ****P < 0.0001). (e) Representative sequential fluorescence images of M. marinum (Wasabi) infected zebrafish embryos following 2 or 4 days of treatment (3 and 5 dpi) with compound 12 at 0.25, 0.5, and 2 μg/mL, relative to untreated zebrafish (UNT). Scale bar, 210 μm. (f) M. marinum infection burden in zebrafish embryo as quantified following 2 (3 dpi) and 4 (5 dpi) days of compound 12 exposure (0.25, 0.5, and 2 μg/mL) by pixel count (fluorescent events) using the ImageJ software. The data presented are the cumulative data of three independent experiment (each containing 20 to 25 embryos per group), with each data point representing one infected zebrafish larva. The error bar represents the mean and standard deviations of the cumulative dataset. Statistical comparison of the different groups was performed using a Mann–Whitney’s t test, (***P < 0.001, ****P < 0.0001).