Fig. 4

Fig. 4. Inhibition of Lipid metabolism via NDGA alters macrophage behavioural switch (A) The biosynthetic pathways of SPM leading to lipoxins, D-series resolvins and Protectins. Lipoxins (LXA4) are derived from arachidonic acid (AA) through the action of 15-Lipoxygenase (15-LOX) and Lipoxygenase 5 (5-LOX). D-series resolvins (RvD1 … 6) and protectins (PD1) are derived from docosahexaenoic acid (DHA) by the action 15-LOX and 5-LOX and from 15-LOX only, respectively. (B) Schedule of the experiment. The fin folds of Tg(mfap4:mcherry-F) larvae were amputated at 3 dpf and immediately treated with DMSO or NDGA 50 μM. Larvae were imaged in live at 6 and 13 hpA. (C) Representative images of the fin folds from amputated Tg(mfap4:mcherry-F) larvae at 13 hpA after DMSO or NDGA 50 μM treatment. Images are maximum projections from confocal microscopy showing the fluorescence of mCherry-F (macrophages) in the fin folds. White lines outline the fin fold. Scale bar: 100 μm. (C′) Zoomed region of the wound at 13 hpA after NDGA 50 μM treatment. Maximum projection from confocal microscopy z-stack showing the fluorescence of mCherry-F (macrophages) overlaid on a brightfield image. Arrow heads show macrophages around pigments. Scale bar: 100 μm (D) Quantification of macrophage distance from the wound (values in μm) in DMSO or NDGA -treated larvae at 6 and 13 hpA. Mean±sem, three independent experiments, the number of analysed macrophages in parenthesis, kruskal-Wallis test with Dunn's post-test, ***p < 0.001, ns – not significant. (E) Quantification of macrophage perimeter in DMSO or NDGA -treated larvae at 6 and 13 hpA. Mean±sem, three independent experiments, the number of analysed macrophages is indicated in parenthesis, kruskal-Wallis test with Dunn's post-test, **p < 0.01, ***p < 0.001. (F) Quantification of macrophage circularity in DMSO or NDGA -treated larvae at 6 and 13 hpA. Mean±sem, three independent experiments, the number of analysed macrophages is indicated in parenthesis, kruskal-Wallis test with Dunn's post-test, ***p < 0.001.