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Fig. 2
In Vivo Phagolysosomal Trafficking of M. marinum
(A) Illustration of zebrafish larva with injection sites outlined in red. The hindbrain ventricle (HBV) is accessible to recruited myeloid cells, while the caudal vein (CV) traverses the caudal hematopoietic tissue where myeloid cells develop.
(B) Confocal images of blue fluorescent Mm that have been phagocytosed by green fluorescent macrophages in the brain of a 2 day post-fertilization (dpf) larva stained with red LT; one bacterium shown colocalizes with LT, and the other does not. Scale bar, 10 µm.
(C) Percent of Mm colocalizing with LT dye 24 hr post infection (hpf) into the HBV, representative of three experiments.
(D) Percent of Mm colocalizing with LT dye at 3, 8, and 24 hpi in the CV, representative of three experiments.
(E) Confocal images of blue fluorescent Mm that were pre-labeled with red pHrodo prior to infection into the HBV of 2dpf larvae with green fluorescent macrophages; bacteria that colocalize with pHrodo (arrowheads) and one that does not (arrow). Scale bar, 10 µm.
(F) Percent of Mm colocalizing with pHrodo at 3 and 24 hpi in the HBV or CV of a 3 dpf larva, representative of two experiments.
(G and H) Percent of Mm colocalizing with DQ-BSA (G) or MR-Cathepsin (MRC) (H) imaged at 24 hpi following infection in the HBV or CV of 2 dpf larvae, representative of two experiments each.
(I) Percent of live, heat-killed, or ptpA::Tn Mm pre-labeled with pHrodo prior to infection into the HBV of 2 dpf larvae imaged at 24 hpi, representative of three experiments.
(J) Percent of ΔESX-1 Mm colocalizing with pHrodo imaged at 24 hpi in the HBV of 2 dpf larvae, representative of three experiments. Significance tested using one-way ANOVA with Tukey′s post-test (F and I) or two-tailed unpaired t test (G, H, and J). Each point in (C), (D), and (F)-(J) represents one larva, with mean depicted as a horizontal line. See also Figures S1 and S2.