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Fig. 4
Lysosomal Trafficking Fails to Eradicate Mm Infection
(A) Cartoon diagram of infectivity experiment. Scale bar, 10 µm.
(B) Percent of zebrafish larvae still infected at 60 hpi following infection in the HBV with single pHrodo-labeled Mm and sorting into acidified/non-acidified groups, representative of three experiments.
(C) Photobleaching assay to discern live from killed bacteria at the end of the infectivity assay showing a live bacterium that recovers fluorescence and a dead one that does not. Scale bar, 5 µm.
(D) Data in (B) amended so that larvae with only non-recovering bacteria are placed into the uninfected category, representative of three experiments.
(E) Bacterial numbers at the end of the infectivity experiment, showing only bacteria that recovered after photobleaching, representative of three experiments. p values in parentheses reflect the statistical significance of comparing each larva in that group to the starting bacterial number (1 in each fish) in a Wilcoxon matched-pair signed rank test.
(F-H) Percentage of zebrafish larvae still infected at 60 hpi which contained pHrodo-labeled marP::Tn (F), ptpA::Tn (G), and ΔESX-1 (H) Mm separated by whether the bacteria were pHrodo-positive (lysosomal) or pHrodo-negative (phagosomal) at sorting, representative of two (ΔESX-1) or three (marP::Tn, ptpA::Tn) experiments. Significance tested using two-tailed unpaired t test (E) or Fisher’s exact test (B, D, and F-H). Each point in (E) represents one larva. See also Figure S4.