Fig. 7

Recruitment of neutrophils is crucial for granuloma development.

(A and B) WT or tnfr morphants were iv infected with R Mabs (tdTomato, ≈150 CFU). Kinetics of granuloma formation (A) and mean ± SEM number of granuloma per infected embryos (B) in (A); (n = 30–45, three independent experiments). The top panel shows confocal images of representative granuloma. Scale bars, 100 μm. (C) Confocal images showing a representative 3 dpi granulomas in WT versus tnfr1 morphant Tg(mpeg:mcherry-F) embryos iv infected with S expressing Wasabi. Arrows indicate extracellular aggregates. Scale bars, 15 μm. (D) Confocal microscopy showing representative granulomas in WT and tnfr1 morphant Tg(mpx:eGFP) embryos iv infected with R expressing tdTomato at 4 dpi. Arrows indicate extracellular cords. Scale bars, 30 μm. (E-F) Confocal microscopy monitored WT or tnfr morphant Tg(mpx:eGFP) embryos that were iv infected with R (tdTomato, ≈150 CFU). (E) Representative kinetics of neutrophil mobilization during granuloma formation. Scale bars, 20 μm. (F) Number of neutrophils recruited to WT (up) or TNFR1-depleted (down) granuloma as a function of granuloma volume. Statistical significance was determined by Fisher’s exact test of a contingency table (A), one-tailed unpaired Student’s t test (B) or Pearson correlation (F). See also S6 and S7 Movies.