Figure 3.

miR-126 potentially targets tsc1a to worsen M. marinum infection burden.

(A) Binding kinetics of tsc1a and dre-miR-126a-3p as predicted by RNAhybrid. (B) Brightfield images of tsc1a knockdown embryos at 3 dpf showing no abnormal developmental phenotypes. Scale bar represents 200 μm. (C)tsc1a expression was measured by RT-qPCR at 1 and 3 dpi after CRISPR-Cas9 knockdown of tsc1a and infection with M. marinum. (D)tsc1a knockdown and scramble control embryos were infected with M. marinum via caudal vein injection, and the bacteria burden was analysed at 1 and 3 dpi. (E)tsc1a and miR-126 double knockdown embryos were infected with M. marinum via caudal vein injection, and bacterial burden was analysed at 3 dpi. Data information: for RT-qPCR analysis, data points are representative of a single measurement of 10 pooled embryos and two technical replicates, with the mean and SEM shown and comparisons calculated by one-way ANOVA. (C, D) Bacterial burden data points represent a single measurement (n = 27–44 embryos per group (C) and n = 11–20 embryos per group (D)), and two experimental replicates with the mean and SEM are shown and comparisons calculated by one-way ANOVA.