Fig. S4

Ablation of TNF signaling does not affect the early chemoattraction of macrophages to localized Mabs infections.

To evaluate the effect of absence of TNF signaling on early and late macrophages recruitment, WT or tnfr morphants Tg(mpeg1:mCherry-F) larvae were injected with either PBS or Wasabi-expressing Mabs (R variant) into the muscle (A) or otic cavity (B-C), monitored and imaged using confocal microscopy to measure macrophage recruitment. (A) Representative confocal microscopy of macrophages recruitment into the infected muscle at 2 hpi (dotted line outlines 2 somitic muscles). Scale bars, 100 μm. (B-C) Dynamic of macrophage recruitment at the infection site assessed at 2, 4 and 6 hpi (Scale bars, 50 μm) (B) and number of recruited macrophages at 2 hpi (C). Results are presented as average number from two experiments. Each symbol represents individual embryos and horizontal lines indicate mean values. Significance was assessed by one-tailed unpaired Student’s t test. In both WT- and tnfr morphant-infected animals, macrophages are recruited towards bacteria at the same rate at early time post-infection. However, while the number of newly recruited macrophages increased progressively in WT larvae from 2 hpi to 6 hpi, the number of recruited macrophages remains constant in tnfr morphants.