Fig. 1
Binary transgenic zebrafish model for regulatory profiling of neutrophils in response to M. marinum. (A) Driver line expressing BirA in neutrophils (i) is crossed with effector line ubiquitously expressing Avi-tagged Rangap (ii) for biotinylation of nuclei. In double transgenic fish, only the nuclei of neutrophils will be biotinylated (iii). (B) Effector line with ubiquitously expressed Avi-Cerulean-Rangap. (C) Neutrophil-specific BirA driver line with Citrine reporter. (D) Time course analysis of neutrophil infiltration of M. marinum. Embryos were injected with M. marinum at the 32-512-cell stage, fixed and immunofluorescence carried out for mpx protein at 2 dpi (i), 3 dpi (ii), 4 dpi (iii) and 5 dpi (iv). Neutrophil localisation is indicated by white arrows. (E) Neutrophil numbers in proximity to M. marinum were quantified at 2–5 dpi. Statistical significance was determined by one way ANOVA (n = 14–18, ****p < 0.0001). (F) Infection model for subsequent experiments.