Fig 8

(A–C) Three examples of in vitro shuttling of zymosan–pHrodo particles, representative of 164 shuttling events. Neutrophils, green; macrophages, red; pHrodo–zymosan, false-colored blue. Scales as shown. Stills from S6C–S6E Movie. (D) Distribution of the time of shuttling for 164 in vitro shuttling events in panel C and S2 Table (Experiment 1, n = 66; Experiment 2, n = 98). (E) pHrodo signal intensity (arbitrary units) for in vitro shuttled zymosan–pHrodo particles (n = 164). Background intensity is for n = 100 randomly selected points clearly positioned between cells (n = 50 from each experiment). Black dots show the distributions of zymosan–pHrodo intensities in neutrophils 6 min before shuttling and in macrophages 6 min after shuttling; red lines connect paired values. p-values from unpaired two-tailed t test (background versus neutrophil) and paired two-tailed t test (neutrophil versus macrophage). Green line indicates median. Blue lines (1–3) correspond to the 3 examples shown in panels A–C. (F) Example of in vitro shuttling of a zymosan–pHrodo particle from a wild-type donor neutrophil to a Dectin-1^−/− recipient macrophage. Scale as shown. Stills from S6F Movie. Colored arrowheads (in A–C and D) indicate the conidium within donor neutrophil (green), at the point of intercellular transfer (white), and in the recipient macrophage (red). Datasets for D–E are provided in S1 Data. Exp, experiment; WT, wild type.